Methods for the detection, visualization and high resolution physical mapping of genomic rearrangements in breast and ovarian cancer genes and loci brca1 and brca2 using genomic morse code in conjunction with molecular combing

ABSTRACT

Methods for detecting genomic rearrangements in BRCA1 and BRCA2 genes at high resolution using Molecular Combing and for determining a predisposition to a disease or disorder associated with these rearrangements including predisposition to ovarian cancer or breast cancer. Primers useful for producing probes for this method and kits for practicing the methods.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of U.S. Ser. No. 13/665,404, filed Oct. 31, 2012, which claims priority to U.S. Provisional Application No. 61/553,906, filed Oct. 31, 2011, the entire contents of which are incorporated herein by reference. On Oct. 30, 2012, International Application PCT/IB/2012/002422 was also filed with the same title, the entire contents of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to a method for detecting genomic rearrangements in BRCA1 and BRCA2 genes and loci at high resolution using Molecular Combing and relates to a method of determining a predisposition to diseases or disorders associated with these rearrangements including predisposition to ovarian cancer or breast cancer.

2. Description of the Related Art

Breast cancer is the most common malignancy in women, affecting approximately 10% of the female population. Incidence rates are increasing annually and it is estimated that about 1.4 million women will be diagnosed with breast cancer annually worldwide and about 460,000 will die from the disease. Germline mutations in the hereditary breast and ovarian cancer susceptibility genes BRCA1 (MIM#113705) and BRCA2 (MIM#600185) are highly penetrant (King et al., 2003), (Nathanson et al., 2001). Screening is important for genetic counseling of individuals with a positive family history and for early diagnosis or prevention in mutation carriers. When a BRCA1 or BRCA2 mutation is identified, predictive testing is offered to all family members older than 18 years. If a woman tests negative, her risk becomes again the risk of the general population. If she tests positive, a personalized surveillance protocol is proposed:

it includes mammographic screening from an early age, and possibly prophylactic surgery. Chemoprevention of breast cancer with anti-estrogens is also currently tested in clinical trial and may be prescribed in the future.

Most deleterious mutations consist of either small frameshifts (insertions or deletions) or point mutations that give rise to premature stop codons, missense mutations in conserved domains, or splice-site mutations resulting in aberrant transcript processing (Szabo et al., 2000). However, mutations also include more complex rearrangements, including deletions and duplications of large genomic regions that escape detection by traditional PCR-based mutation screening combined with DNA sequencing (Mazoyer, 2005).

Techniques capable of detecting these complex rearrangements include Southern blot analysis combined with long-range PCR or the protein truncation test (PTT), quantitative multiplex PCR of short fluorescent fragments (QMPSF) (Hofmann et al., 2002), real-time PCR, fluorescent DNA microarray assays, multiplex ligation-dependent probe amplification (MLPA)(Casilli et al., 2002), (Hofmann et al., 2002) and high-resolution oligonucleotide array comparative genomic hybridization (aCGH) (Rouleau et al., 2007), (Staaf et al., 2008). New approaches that provide both prescreening and quantitative information, such as qPCR-HRM and EMMA, have recently been developed and genomic capture combined with massively parallel sequencing has been proposed for simultaneous detection of small mutations and large rearrangements affecting 21 genes involved in breast and ovarian cancer (Walsh et al., 2010).

Molecular Combing is a powerful FISH-based technique for direct visualization of single DNA molecules that are attached, uniformly and irreversibly, to specially treated glass surfaces (Herrick and Bensimon, 2009); (Schurra and Bensimon, 2009). This technology considerably improves the structural and functional analysis of DNA across the genome and is capable of visualizing the entire genome at high resolution (in the kb range) in a single analysis. Molecular Combing is particularly suited to the detection of genomic imbalances such as mosaicism, loss of heterozygosity (LOH), copy number variations (CNV), and complex rearrangements such as translocations and inversions (Caburet et al., 2005), thus extending the spectrum of mutations potentially detectable in breast cancer genes. Molecular Combing has been successfully employed for the detection of large rearrangements in BRCA1 ((Gad et al., 2001), (Gad et al., 2002a), (Gad et al., 2003) and BRCA2 (Gad et al., 2002b), using a first-generation “color bar coding” screening approach. However, these techniques lack resolution and cannot precisely detect large rearrangements in and around BRCA1 and BRCA2.

In distinction to the prior art techniques, as disclosed herein, the inventors provide a novel Genetic Morse Code Molecular Combing procedure that provides for high resolution visual inspection of genomic DNA samples, precise mapping of mutated exons, precise measurement of mutation size with robust statistics, simultaneous detection of BRCA1 and BRCA2 genetic structures or rearrangements, detection of genetic inversions or translocations, and substantial elimination of problems associated with repetitive DNA sequences such as Alu sequences in BRCA1 and BRCA2 loci.

BRIEF SUMMARY OF THE INVENTION

The BRCA1 and BRCA2 genes are involved, with high penetrance, in breast and ovarian cancer susceptibility. About 2% to 4% of breast cancer patients with a positive family history who are negative for BRCA1 and BRCA2 point mutations can be expected to carry large genomic alterations (deletion or duplication) in one of the two genes, and especially BRCA1. However, large rearrangements are missed by direct sequencing. Molecular Combing is a powerful FISH-based technique for direct visualization of single DNA molecules, allowing the entire genome to be examined at high resolution in a single analysis. A novel predictive genetic test based on Molecular Combing is disclosed herein. For that purpose, specific BRCA1 and BRCA2 “Genomic Morse Codes” (GMC) were designed, covering coding and non-coding regions and including large genomic portions flanking both genes. The GMC is a series of colored signals distributed along a specific portion of the genomic DNA which signals arise from probe hybridization with the probes of the invention. The concept behind the GMC has been previously defined in WIPO patent application WO/2008/028931 (which is incorporated by reference), and relates to the method of detection of the presence of at least one domain of interest on a macromolecule to test.

A measurement strategy is disclosed for the GMC signals, and has been validated by testing 6 breast cancer patients with a positive family history and 10 control patients. Large rearrangements, corresponding to deletions and duplications of one or several exons and with sizes ranging from 3 kb to 40 kb, were detected on both genes (BRCA1 and BRCA2). Importantly, the developed GMC allowed to unambiguously localize several tandem repeat duplications on both genes, and to precisely map large rearrangements in the problematic Alu-rich 5′-region of BRCA1. This new developed Molecular Combing genetic test is a valuable tool for the screening of large rearrangements in BRCA1 and BRCA2 and can optionally be combined in clinical settings with an assay that allows the detection of point mutations.

A substantial technical improvement compared to the prior color bar coding approach is disclosed here that is based on the design of second-generation high-resolution BRCA1 and BRCA2 Genomic Morse Codes (GMC). Importantly, repetitive sequences were eliminated from the DNA probes, thus reducing background noise and permitting robust measurement of the color signal lengths within the GMC. Both GMC were statistically validated on samples from 10 healthy controls and then tested on six breast cancer patients with a positive family history of breast cancer. Large rearrangements were detected, with a resolution similar to the one obtained with a CGH (1-3 kb). The detected mutation demonstrates the robustness of this technology, even for the detection of problematic mutations, such as tandem repeat duplications or mutations located in genomic regions rich of repetitive elements. The developed Molecular Combing platform permits simultaneous detection of large rearrangements in BRCA1 and BRCA2, and provides novel genetic tests and test kits for breast and ovarian cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color.

FIGS. 1A and 1B: Dot plot alignments of the human BRCA1 and BRCA2 genomic regions. Dot plot matrix showing self-alignment of the 207-kb genomic regions derived from the BAC RP11-831F13 (ch17:41172482-41379594) encoding BRCA1 (1A), and the 172-kb genomic regions derived from the BAC RP11-486017 (ch13: 32858070-33030569) encoding BRCA2 (1B), based on the GRCh37 genome assembly (also called hg19, April 2009 release) and using JDotter software (URL:http://_athena.bioc.uvic.ca/tools/JDotter). The main diagonal represents alignment of the sequence with itself, while the lines out of the main diagonal represent similar or repetitive patterns within the sequence. The dark regions contain large numbers of repetitive sequences, whereas the bright regions contain none. The genes are represented as arrows in the 5′→3′ direction. The sizes and BAC coordinates of the genomic regions, encoding for repetitive sequences, not included in the DNA probes are indicated in the tables on the left. The bottom panels indicate the name and the size (in kb) of the DNA probes (35 for BRCA1 and 27 for BRCA2) without potentially disturbing repetitive sequences, derived from the bioinformatics analysis.

FIGS. 2A, 2B, 2C and 2D: In silico-generated Genomic Morse Codes designed for high-resolution physical mapping of the BRCA1 and BRCA2 genomic regions. Probes colors are represented here as grayscale variations: blue probes are shown as black boxes, green probes as white boxes and red probes as gray boxes. (2A) The complete BRCA1 GMC covers a genomic region of 200 kb and is composed of 18 signals (S1B1-S18B) of a distinct color (green, red or blue). Each signal is composed of 1 (e.g., S2B1) to 3 small horizontal bars (e.g., S15B1), each bar corresponding to a single DNA probe. The region encoding the BRCA1 gene (81.2 kb) is composed of 7 “motifs” (g1b1-g7b1). Each motif is composed of 1 to 3 small horizontal bars and a black “gap” (no signal). (2B) Zoom-in on the BRCA1 gene-specific signals and relative positions of the exons. (2C) The complete BRCA2 GMC covers a genomic region of 172 kb and is composed of 14 signals (S1B2-S14B2) of a distinct color (green, red or blue). Each signal is composed of 1 (e.g., S14B2) to 5 small horizontal bars (e.g., S1B2). The region encoding the BRCA2 gene (84.2 kb) is composed of 5 motifs 24 (g1b2-g5b2). Each motif is composed of 2 to 4 small horizontal bars and a black gap. (2D) Zoom-in on the BRCA2 gene-specific signals and relative positions of the exons. Deletions or insertions, if present, will appear in the region covered by the motifs.

FIGS. 3A and 3B: Validation of BRCA1 and BRCA2 Genomic Morse Code signals in control patients. Original microscopy images consist of three channel images where each channel is the signal from a given fluorophore—these are acquired separately in the microscopy procedure. These channels are represented here as different shades on a grayscale: blue probes are shown in black, green probes in white and red probes in dark gray, while background (absence of signal) is light gray. In diagrams, the same convention as in FIG. 2 is used. The aspect ratio was not preserved, signals have been “widened” (i.e. stretched perpendicularly to the direction of the DNA fiber) in order to improve the visibility of the probes. Typical BRCA1 (3A) and BRCA2 (3B) Genomic Morse Code signals and measured motif lengths (kb) in one control patient (absence of large rearrangements) are reported. The BRCA1 and BRCA2 signals obtained after microscopic visualization are shown at the top of the tables, including the position of the motifs related to the gene of interest. Typically 20 to 40 images (n^(o) images) were selected, and motifs were measured with GVLab software. For each motif, the following values were determined: the theoretical calculated length (calculated (kb)), the mean measured length (μ (kb)), the standard deviation (SD (kb)), the coefficient of variation (CV (%)), the difference between μ and calculated (delta), and the stretching factor (SF=(calculated/μ)×2). In the absence of mutations, SF values are comprised between 1.8 and 2.2 and delta values are comprised between −1.9 kb and 1.9 kb (see Material and Methods in Example 1 for details).

FIGS. 4A, 4B, and 4C: Known BRCA1 large rearrangements detected in breast cancer patients.

As in FIGS. 2 and 3, diagrams and microscopy images are represented in shades of gray, with the following correspondence: blue is shown as black, green as white and red as dark gray (on a light gray background) and aspect ratio in microscopy images may have been modified for clarity. DNA isolated from EBV-immortalized B lymphocytes collected from breast cancer patients was analyzed by Molecular Combing to confirm known large rearrangements previously characterized by aCGH (see Table 3). Three large rearrangements out of seven are shown in the figure: (4A) Dup ex 13 (case 1), visible as a tandem repeat duplication of the blue signal S7B1. The g4B1 motif (16.5 kb) was first measured on a mixed population of 40 images, comprising wild type and mutated alleles, and following values were obtained: μ(BRCA1^(wt)+BRCA1^(mt) signals)=19 kb±3.5 kb, delta=2.5 kb (duplication is confirmed since delta≧2 kb). The images were then divided in two groups: 21 images were classified as BRCA1^(wt), and 19 images were classified as BRCA1^(mt). The size was then calculated as the difference between the motif mean sizes of the two alleles: μ(BRCA1^(wt))=16.1±1.6 kb, μ(BRCA1^(mt))=22.2±2.0 kb, mutation size=μ(BRCA1^(mt))−μ(BRCA1^(wt))=6.1±1.6 kb. The bottom panel shows the MLPA fragment display (left) and the normalized MLPA results (right), arrows indicating exons interpreted as duplicated. (4B) Del ex 8-13 (case 6), visible as a deletion of the blue signal S7B1, including a large genomic portion between signals S7B1 and S8B1. The g4B1 (16.5 kb) and the g5b1 (19.7 kb) motifs were first measured on a mixed population of 23 images, yielding following values. For g4b1: μ(BRCA1^(wt)+BRCA1^(mt))=17.5±4.0 kb, delta=−2.2 kb (delta≦−2 kb); 13 images were then classified as BRCA1^(wt) and 10 images as BRCA μ(BRCA1^(wt))=20.8±1.6 kb, μ(BRCA1^(mt))=13.3±1.1 kb, μ(BRCA1^(mt))−μ(BRCA1^(wt))=−7.5±1.6 kb. For g5b1: μ(BRCA1^(wt)+BRCA1^(mt))=12.8±5.5 kb, delta=−3.7 kb (delta≦−2 kb); 13 images were then classified as BRCA1^(wt) and 10 images as BRCA1^(mt): μ(BRCA1^(wt))=18.3±1.3 kb, μ(BRCA1^(mt))=5.8±0.5 kb, μ(BRCA1^(mt))−μ(BRCA1^(wt))=−12.5±1.0 kb. Total mutation size=mutation size g4B1+mutation size g5b1=−20±2.8 kb. (4C) Del ex 2 (case 2), visible as a deletion of the green signal S10B1, as well as a large genomic portion of the 5′ region upstream of BRCA1, including S11B1 and S12B1. To confirm the presence of the deletion in the BRCA1 gene, the g7B1 (17.7 kb) motif was first measured on a mixed population of 20 images, yielding following values: μ(BRCA1^(wt)+BRCA1^(mt))=12.3±2.9 kb, delta=−5.4 kb (deletion is confirmed since delta≦−2 kb). To measure mutations size within the BRCA1 gene, 11 images were then classified as BRCA1^(wt) and 9 images as BRCA1^(mt), yielding following values: μ(BRCA1^(wt))=18.1±0.7 kb, μ(BRCA1^(mt))=8.1±1.6 kb, mutation size=μ(BRCA1^(mt))−μ(BRCA1^(wt))=−10±1.5 kb. To include the deleted genomic region upstream of BRCA1 and determine the whole mutation size, we had to measure the genomic region between the signals S8B1 and S14B1 (89.9 kb). The S8B1-S14B1 region was first measured on 19 images, yielding following values: μ(BRCA1^(wt)+BRCA1^(mt))=62.3±18.4 kb, delta=−27.6 kb. 11 images were then classified as BRCA1^(wt), and 8 images as BRCA1^(mt), yielding following values: μ(BRCA1^(wt))=92.2±3.2 kb, μ(BRCA1^(mt))=51.4±2.2 kb, mutation size=μ(BRCA1^(mt))−μ(BRCA1^(wt))=−40.8±3.5 kb. The BRCA1 signals, derived from both the wild-type (=BRCA1^(wt)) and the mutated allele (=BRCA1^(mt)), obtained after microscopic visualization, are shown in the top panels. The position, nature (deletion or duplication) and size (in kb) of the detected large rearrangements are indicated in orange. The zoom-in on the BRCA1 gene-specific signals and the relative positions of the mutated exons are shown in the bottom panels. mt, mutated allele; wt, wild-type allele.

FIG. 5. GMC used for BRCA1. Another example of a high resolution genomic morse code to analyze the BRCA1 gene region is shown here. As in FIG. 2, diagrams are represented with the following correspondence: blue probes are shown as black, green as white and red as dark gray.

FIG. 6: Duplication in exons 18-20 of BRCA1

The GMC described in FIG. 2, with probe labels modified as shown in the diagram, was hybridized on this sample. As in FIGS. 2 and 3, diagrams and microscopy images are represented in shades of gray, with the following correspondence: blue is shown as black, green as white and red as dark gray (on a light gray background) and aspect ratio in microscopy images may have been modified for clarity. By visual inspection, there appears to be a tandem duplication of the red signal S5B1. After measurement, the mutation was estimated to have a size of 6.7±1.2 kb, restricted to a portion of the genome that encodes for exons 18 to 20. The estimated mutation size is fully in line with the 8.7 kb reported in the literature (Staaf, 2008). Details on the measurement and statistical analysis can be found in Example 1.

FIG. 7 9: examples of Alu sequences excluded from the BRCA1 (A) and BRCA2 (B) GMCs.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Physical mapping: is the creation of a genetic map defining the position of particular elements, mutations or markers on genomic DNA, employing molecular biology techniques. Physical mapping does not require previous sequencing of the analyzed genomic DNA.

FISH: Fluorescent in situ hybridization.

Molecular Combing: a FISH-based technique for direct visualization of single DNA molecules that are attached, uniformly and irreversibly, to specially treated glass surfaces.

Predictive genetic testing: screening procedure involving direct analysis of DNA molecules isolated from human biological samples (e.g.: blood), used to detect gene mutations associated with disorders that appear after birth, often later in life. These tests can be helpful to people who have a family member with a genetic disorder, but who have no features of the disorder themselves at the time of testing. Predictive testing can identify mutations that increase a person's chances of developing disorders with a genetic basis, such as certain types of cancer.

Polynucleotides: This term encompasses naturally occurring DNA and RNA polynucleotide molecules (also designated as sequences) as well as DNA or RNA analogs with modified structure, for example, that increases their stability. Genomic DNA used for Molecular Combing will generally be in an unmodified form as isolated from a biological sample. Polynucleotides, generally DNA, used as primers may be unmodified or modified, but will be in a form suitable for use in amplifying DNA. Similarly, polynucleotides used as probes may be unmodified or modified polynucleotides capable of binding to a complementary target sequence. This term encompasses polynucleotides that are fragments of other polynucleotides such as fragments having 5, 10, 15, 20, 30, 40, 50, 75, 100, 200 or more contiguous nucleotides.

BRCA1 locus: This locus encompasses the coding portion of the human BRCA1 gene (gene ID: 672, Reference Sequence NM_(—)007294) located on the long (q) arm of chromosome 17 at band 21, from base pair 41,196,311 to base pair 41,277,499, with a size of 81 kb (reference genome Build GRCh37/hg19), as well as its introns and flanking sequences. Following flanking sequences have been included in the BRCA1 GMC: the 102 kb upstream of the BRCA1 gene (from 41,277,500 to 41,379,500) and the 24 kb downstream of the BRCA1 gene (from 41,196,310 to 41,172,310). Thus the BRCA1 GMC covers a genomic region of 207 kb.

BRCA2 locus: This locus encompasses the coding portion of the human BRCA2 gene (gene ID: 675, Reference Sequence NM_(—)000059.3) located on the long (q) arm of chromosome 13 at position 12.3 (13q12.3), from base pair 32,889,617 to base pair 32,973,809, with a size of 84 kb (reference genome Build GRCh37/hg19), as well as its introns and flanking sequences. Following flanking sequences have been included in the BRCA2 GMC: the 32 kb upstream of the BRCA2 gene (from 32,857,616 to 32,889,616) and the 56 kb downstream of the BRCA2 gene (from 32,973,810 to 33,029,810). Thus the BRCA2 GMC covers a genomic region of 172 kb.

Germline rearrangements: genetic mutations involving gene rearrangements occurring in any biological cells that give rise to the gametes of an organism that reproduces sexually, to be distinguished from somatic rearrangements occurring in somatic cells.

Point mutations: genetic mutations that cause the replacement of a single base nucleotide with another nucleotide of the genetic material, DNA or RNA. Often the term point mutation also includes insertions or deletions of a single base pair.

Frameshift mutations: genetic mutations caused by indels (insertions or deletions) of a number of nucleotides that is not evenly divisible by three from a DNA sequence. Due to the triplet nature of gene expression by codons, the insertion or deletion can change the reading frame (the grouping of the codons), resulting in a completely different translation from the original.

Tandem repeats duplications: mutations characterized by a stretch of DNA that is duplicated to produce two or more adjacent copies, resulting in tandem repeats.

Tandem repeat array: a stretch of DNA consisting of two or more adjacent copies of a sequence resulting in gene amplification. A single copy of this sequence in the repeat array is called a repeat unit. Gene amplifications occurring naturally are usually not completely conservative, i.e. in particular the extremities of the repeated units may be rearranged, mutated and/or truncated. In the present invention, two or more adjacent sequences with more than 90% homology are considered a repeat array consisting of equivalent repeat units. Unless otherwise specified, no assumptions are made on the orientation of the repeat units within a tandem repeat array.

Complex Rearrangements: any gene rearrangement that can be distinguished from simple deletions or duplications. Examples are translocations or inversions.

Probe: This term is used in its usual sense for a polynucleotide of the invention that hybridizes to a complementary polynucleotide sequences (target) and thus serves to identify the complementary sequence. Generally, a probe will be tagged with a marker, such as a chemical or radioactive market that permits it to be detected once bound to its complement. The probes described herein are generally tagged with a visual marker, such as a fluorescent dye having a particular color such as blue, green or red dyes. Probes according to the invention are selected to recognize particular portions or segments of BRCA1 or BRCA2, their exons or flanking sequences. For BRCA1, probes generally range in length between 200 bp and 5,000 bp. For BRCA2, probes generally range in length between 200 bp and 6,000 bp. The name and the size of probes of the invention are described in FIG. 2. Representative probes according to the invention, such as BRCA1-1A (3,458 bp) or BRCA2-1 (2,450 bp), are described in Tables 1 and 2. In a particular embodiment of the invention, the probes are said to be “free of repetitive nucleotidic sequences”. Such probes may be located in genomic regions of interest which are devoid of repetitive sequences as defined herein.

Detectable label or marker: any molecule that can be attached to a polynucleotide and which position can be determined by means such as fluorescent microscopy, enzyme detection, radioactivity, etc, or described in the US application nr. US2010/0041036A1 published on 18 Feb. 2010.

Primer: This term has its conventional meaning as a nucleic acid molecule (also designated sequence) that serves as a starting point for polynucleotide synthesis. In particular, Primers may have 20 to 40 nucleotides in length and may comprise nucleotides which do not base pair with the target, providing sufficient nucleotides in their 3′-end, especially at least 20, hybridize with said target. The primers of the invention which are described herein are used to produce probes for BRCA1 or BRCA2, for example, a pair of primers is used to produce a PCR amplicon from a bacterial artificial chromosome as template DNA. The sequences of the primers used herein are referenced as SEQ ID 1 to SEQ ID 130 in Table 8. In some cases (details in table 1), the primers contained additional sequences to these at their 5′ end for ease of cloning. These additional sequences are SEQ ID 134 (containing a poly-A and a restriction site for AscI) for forward primers and SEQ ID 135 (containing a poly-A and a restriction site for PacI) for reverse primers.

Tables 1 and 2 and 8 describe representative primer sequences and the corresponding probe coordinates.

Genomic Morse Code(s): A GMC is a series of “dots” (DNA probes with specific sizes and colors) and “dashes” (uncolored spaces with specific sizes located between the DNA probes), designed to physically map a particular genomic region. The GMC of a specific gene or locus is characterized by a unique colored “signature” that can be distinguished from the signals derived by the GMCs of other genes or loci. The design of DNA probes for high resolution GMC requires specific bioinformatics analysis and the physical cloning of the genomic regions of interest in plasmid vectors. Low resolution CBC has been established without any bioinformatics analysis or cloning procedure.

Repetitive nucleotidic sequences: the BRCA1 and BRCA2 gene loci contain repetitive sequences of different types: SINE, LINE, LTR and Alu. The repetitive sequences which are present in high quantity in the genome sequence but are absent from the probes, i.e. were removed from the BRCA1 and BRCA2 GMCs of the invention, are mainly Alu sequences, having lengths of about 300 bp (see Figure S1, S1, S2 and S3 for more details). This mainly means that the percentage of the remaining Alu-sequences within the DNA probes compared to percentage present in the reference genome is less than 10% and preferably less than 2%. Accordingly, a polynucleotide is said to be “free of repetitive nucleotidic sequences” when at least one type of repetitive sequences (e.g., Alu, SINE, LINE or LTR) selected from the types of repetitive sequences cited above is not contained in the considered probe, meaning that said probes contains less than 10%, preferably less than 2% compared to percentage present in the reference genome. Examples of Alu repeats found in the BRCA1 and 2 genes are given in FIGS. 7A and 7B, while tables 3 and 4 list the repeats identified by RepeatMasker contained in the BAC clone RP11-831F13 covering the genomic region of BRCA1 (FIG. 7A) or in the BAC clone RP11-486017 covering the genomic region of BRCA2 (FIG. 7B). In both cases, Mu repeats are counted separately in regions where our probes hybridize and in the regions excluded from this probe design.

The term “intragenic large rearrangement” as used herein refers to deletion and duplication events that can be observed in a gene sequence, said sequence comprising in a restricted view introns and exons; and in an extended view introns, exons, the 5′ region of said gene and the 3′ region of said gene. The intragenic large rearrangement can also cover any gain or loss of genomic material with a consequence in the expression of the gene of interest.

The term “locus” as used herein refers to a specific position of a gene or other sequence of interest on a chromosome. For BRCA1 and BRCA2, this term refer to the BRCA1 and BRCA2 genes, the introns and the flanking sequences refer to BRCA1/BRCA2+introns and flanking sequences.

The term “nucleic acid” as used herein means a polymer or molecule composed of nucleotides, e.g., deoxyribonucleotides or ribonucleotides, or compounds produced synthetically such as PNA which can hybridize with naturally occurring nucleic acids in a sequence specific manner analogous to that of two naturally occurring nucleic acids, e.g., can participate in Watson-Crick base pairing interactions. Nucleic acids may be single- or double-stranded or partially duplex.

The terms “ribonucleic acid” and “RNA” as used herein mean a polymer or molecule composed of ribonucleotides.

The terms “deoxyribonucleic acid” and “DNA” as used herein mean a polymer or molecule composed of deoxyribonucleotides.

The term “sample” as used herein relates to a material or mixture of materials, typically, although not necessarily, in fluid form, containing one or more components of interest. For Molecular Combing, the sample will contain genomic DNA from a biological source, for diagnostic applications usually from a patient. The invention concerns means, especially polynucleotides, and methods suitable for in vitro implementation on samples.

The terms “nucleoside” and “nucleotide” are intended to include those moieties that contain not only the known purine and pyrimidine bases, but also other heterocyclic bases that have been modified. Such modifications include methylated purines or pyrimidines, acylated purines or pyrimidines, alkylated riboses or other heterocycles. In addition, the terms “nucleoside” and “nucleotide” include those moieties that contain not only conventional ribose and deoxyribose sugars, but other sugars as well. Modified nucleosides or nucleotides also include modifications on the sugar moiety, e.g., wherein one or more of the hydroxyl groups are replaced with halogen atoms or aliphatic groups, or are functionalized as ethers, amines, or the like.

The term “stringent conditions” as used herein refers to conditions that are compatible to produce binding pairs of nucleic acids, e.g., surface bound and solution phase nucleic acids, of sufficient complementarity to provide for the desired level of specificity in the assay while being less compatible to the formation of binding pairs between binding members of insufficient complementarity to provide for the desired specificity. Stringent assay conditions are the summation or combination (totality) of both hybridization and wash conditions.

A “stringent hybridization” and “stringent hybridization wash conditions” in the context of nucleic acid hybridization (e.g., as required for Molecular Combing or for identifying probes useful for GMC) are sequence dependent, and are different under different experimental parameters. Stringent hybridization conditions that can be used to identify nucleic acids within the scope of the invention can include for example hybridization in a buffer comprising 50% formamide, 5×SSC, and 1% SDS at 42° C., or hybridization in a buffer comprising 5.times.SSC and 1% SDS at 65° C., both with a wash of 0.2×SSC and 0.1% SDS at 65° C. Exemplary stringent hybridization conditions can also include a hybridization in a buffer of 40% formamide, 1M NaCl, and 1% SDS at 37° C., and a wash in 1×SSC at 45° C. Alternatively, hybridization to filter-bound DNA in 0.5 M NaHPO₄, 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at 65° C., and washing in 0.1×SSC/0.1% SDS at 68° C. can be employed. Yet additional stringent hybridization conditions include hybridization at 60° C. or higher and 3×SSC (450 mM sodium chloride/45 mM sodium citrate) or incubation at 42° C. in a solution containing 30% formamide, 1 M NaCl, 0.5% sodium sarcosine, 50 mM MES, pH 6.5. Those of ordinary skill will readily recognize that alternative but comparable hybridization and wash conditions can be utilized to provide conditions of similar stringency.

A probe or primer located in a given genomic locus means a probe or a primer which hybridizes to the sequence in this locus of the human genome. Generally, probes are double stranded and thus contain a strand that is identical to and another that is reverse complementary to the sequence of the given locus. A primer is single stranded and unless otherwise specified or indicated by the context, its sequence is identical to that of the given locus. When specified, the sequence may be reverse complementary to that of the given locus. In certain embodiments, the stringency of the wash conditions that set forth the conditions that determine whether a nucleic acid is specifically hybridized to a surface bound nucleic acid. Wash conditions used to identify nucleic acids may include for example a salt concentration of about 0.02 molar at pH 7 and a temperature of at least about 50° C. or about 55° C. to about 60° C.; or a salt concentration of about 0.15 M NaCl at 72° C. for about 15 minutes; or a salt concentration of about 0.2×SSC at a temperature of at least about 50° C. or about 55° C. to about 60° C. for about 15 to about 20 minutes; or, the hybridization complex is washed twice with a solution with a salt concentration of about 2×SSC containing 0.1% SDS at room temperature for 15 minutes and then washed twice by 0.1×SSC containing 0.1% SDS at 68° C. for 15 minutes; or, equivalent conditions. Stringent conditions for washing can also be for example 0.2×SSC/0.1% SDS at 42° C.

A specific example of stringent assay conditions is rotating hybridization at 65° C. in a salt based hybridization buffer with a total monovalent cation concentration of 1.5 M followed by washes of 0.5×SSC and 0.1×SSC at room temperature.

Stringent assay conditions are hybridization conditions that are at least as stringent as the above representative conditions, where a given set of conditions are considered to be at least as stringent if substantially no additional binding complexes that lack sufficient complementarity to provide for the desired specificity are produced in the given set of conditions as compared to the above specific conditions, where by “substantially no more” is meant less than about 5-fold more, typically less than about 3-fold more. Other stringent hybridization conditions are known in the art and may be employed, as appropriate.

“Sensitivity” describes the ability of an assay to detect the nucleic acid of interest in a sample. For example, an assay has high sensitivity if it can detect a small concentration of the nucleic acid of interest in sample. Conversely, a given assay has low sensitivity if it only detects a large concentration of the nucleic acid of interest in sample. A given assay's sensitivity is dependent on a number of parameters, including specificity of the reagents employed (such as types of labels, types of binding molecules, etc.), assay conditions employed, detection protocols employed, and the like. In the context of Molecular Combing and GMC hybridization, sensitivity of a given assay may be dependent upon one or more of: the nature of the surface immobilized nucleic acids, the nature of the hybridization and wash conditions, the nature of the labeling system, the nature of the detection system, etc.

Design of High-Resolution BRCA1 and BRCA2 Genomic Morse Codes

Molecular Combing has already been used to detect large rearrangements in the BRCA1 and BRCA2 genes, but the hybridization DNA probes originally used were part of a low resolution “color bar coding” screening approach and were composed of cosmids, PACs and long-range PCR products only partially covering the BRCA1 and BRCA2 loci. Of importance, the DNA probes also encoded repetitive sequences particularly abundant at the two loci (Gad et al., 2001), (Gad et al., 2002b). As a consequence, detection of the probes often resulted in the superposition of individual colored signals (e.g., yellow spots resulting from superposition of green and red signals) and in strong background noise, undermining the quality of the images and preventing the development of a robust strategy to measure the signals length. Such a low resolution screening approach did not allow the unambiguous visualization of complex mutations, such as tandem repeat duplications (Schurra and Bensimon, 2009), (Herrick and Bensimon, 2009).

The inventors found that high-resolution Genomic Morse Codes (GMC) that were designed by covering more of the BRCA1 and BRCA2 genomic regions and by removing the disturbing repetitive sequences from the DNA probes resolved the problems associated with the prior color bar coding approach.

To visualize the repetitive sequences, dot-plot alignments of the BAC clones used for DNA probe cloning were first performed, based on the Genome Reference Consortium GRCh37 genome assembly (also called hg19, April 2009 release). Based on Repeat Masker analysis (www._repeatmasker.org), the percentages of Alu repetitive DNA in the BRCA1- and BRCA2-encoding BACs were 35% and 17%, respectively (data not shown). This resulted in a dark dot-plot matrix dense in repetitive sequences for BRCA1 (1.6 Alu sequences per 1 kb of DNA, compared to an average in the human genome of only 0.25 Alu/kb), and a brighter dot-plot matrix for BRCA2 (0.64 Alu/kb of DNA) (FIGS. 1A and 1B).

35 genomic regions in the BRCA1 locus and 27 regions in the BRCA2 locus that had significantly less repetitive sequences were identified and were used to design and clone DNA hybridization probes compatible with the visualization process associated with Molecular Combing. The name, size and color of the DNA hybridization probes, and the exons covered by the probes, are shown in FIG. 1 and listed in Tables 1 (BRCA1) and 2 (BRCA2). Adjacent DNA probes of the same color form a signal. Thus, a Genomic Morse Code is composed of sequences of colored signals distributed along a specific portion of the genomic DNA. Colors were chosen to create unique non-repetitive sequences of signals, which differed between BRCA1 and BRCA2. The sizes and the BAC coordinates of the genomic regions, encoding for repetitive sequences, excluded from the BRCA1/BRCA2 GMC DNA probes are shown in Tables 3 & 4. 257 Alu sequences were excluded from the BRCA1 GMC and 85 Alu sequences were excluded from the BRCA2 GMC. Examples of removed Alu sequences from both GMCs are shown in FIG. 7.

To facilitate Genomic Morse Code recognition and measurement, signals located on the genes were grouped together in specific patterns called “motifs”. An electronic reconstruction of the designed BRCA1 and BRCA2 Genomic Morse Codes is shown in FIG. 2. In this design, the BRCA1 Genomic Morse Code covers a region of 200 kb, including the upstream genes NBR1, NBR2, LOC100133166, and TMEM106A, as well as the pseudogene ψBRCA1. The complete BRCA1 Genomic Morse Code is composed of 18 signals (S1B1-S18B), and the 8 BRCA1-specific signals are grouped together in 7 motifs (g1b1-g7b1) (FIGS. 2 A and B). The BRCA2 Genomic Morse Code covers a genomic region of 172 kb composed of 14 signals (S1B2-S14B2), and the 7 BRCA2-specific signals are grouped together in 5 motifs (g1b2-g5b2) (FIGS. 2C and 2D). Deletions or insertions, if present, are detected in the genomic regions covered by the motifs.

Validation of BRCA1 and BRCA2 Genomic Morse Code Signals in Control Patients

The newly designed Genomic Morse Codes were first validated on genomic DNA isolated from 10 randomly chosen control patients. Typical visualized signals and measured motif lengths for one control donor are reported in FIG. 3, with BRCA1 at the top and BRCA2 at the bottom. For each Genomic Morse Code, 20 to 30 images were typically analyzed by measuring the length of the different motifs (see nr. images in FIG. 3). Importantly, for all the motifs, the measured values were always similar to the calculated values (compare μ and calculated in FIG. 3). The robustness of BRCA1 and BRCA2 signal measurement was determined by calculating the mean of the measured motif lengths in all 10 control patients, and by comparing the mean measured values with the calculated values (see Table S1). For BRCA1, we obtained delta values (difference between μ and calculated) in the range of −0.2 kb and +0.8 kb, whereas BRCA2 delta values were in the range of −0.3 kb and +0.4 kb, underlining the precision of the developed measurement approach and confirming that the resolution of Molecular Combing is around ±1 kb (Michalet et al., 1997). Molecular Combing allows DNA molecules to be stretched uniformly with a physical distance to contour length correlation of 1 equivalent to 2 kb (Michalet et al., 1997). As a consequence, in the absence of large rearrangements, the derived stretching factor (SF) has a value close to 2 kb/□μm (±0.2). This was confirmed in all the analyzed control donors, with SF values in the range of 1.8-2.2 kb/μm (see SF in FIG. 3). Accordingly, in the presence of large rearrangements in both BRCA1 and BRCA2, SF values are expected to be ≧2.3 kb/μm (for deletions) or ≦1.7 kb/μm (for duplications) and the corresponding delta values are expected to be ≧2 kb (for duplications) or ≦−2 kb (for deletions). Importantly, the presence of a large rearrangement is always validated by visual inspection of the corresponding Genomic Morse Code.

Detection of Known BRCA1 Large Rearrangements in Breast Cancer Patients

Molecular Combing was then applied to 6 samples from patients with a severe family history of breast cancer and known to bear large rearrangements either on BRCA1 or BRCA2 (preliminary screening performed by MLPA or QMPSF). Importantly, the Molecular Combing analysis was a blind test, meaning that for each of the patient the identity of the mutation was unknown before the test, since it was revealed to the operator only after having completed the test on all the samples. 6 different large rearrangements were identified (see Table 5). Importantly, all 6 known mutations have been recently characterized by aCGH and break-point sequencing (Rouleau 2007) and were correctly identified and characterized by Molecular Combing. Complete characterization of the 3 most significant known BRCA1 large rearrangements is reported in FIG. 4 and is described here below.

Duplication of Exon 13 (BRCA1)

By visual inspection via Molecular Combing, this mutation appears as a partial tandem duplication of the blue signal S7B1 (FIG. 4A, top panel). After measurement, the mutation was estimated to have a size of 6.1 kb, restricted to a portion of the DNA probe BRCA1-8 that encodes exon 13. The estimated mutation size is fully in line with the 6.1 kb reported in the literature (Puget 1999), and according to the Breast Cancer Information Core database, this mutation belongs to the 10 most frequent mutations in BRCA1 (Szabo 2000). Duplications are difficult to detect with quantitative methods such as MLPA, often giving rise to false-positive signals (Cavalieri 2007, Staaf 2008). The characterized patient was therefore also analyzed by MLPA, and the duplication of exon 13 was confirmed. More importantly, we also detected a duplication of exons 1A+1B (FIG. 4A, bottom panel), but this mutation could not be detected by Molecular Combing (a duplication of exon 13, if present, would yield two distinct S10B1 signals). Therefore, we consider the exon 1A+1B mutation detected by MLPA to be a false-positive signal. The risk of false-positive signals is more limited in Molecular Combing.

Deletion from Exon 8 to Exon 13 (BRCA1)

By visual inspection, the mutation appeared as a visible as a deletion of the blue signal S7B1, including a large genomic portion between signals S7B1 and S8B1 (FIG. 4B). After measurement, the mutation was estimated to have a size of 26.7 kb in a portion of the BRCA1 gene that encodes from exon 8 to exon 13. The size reported in the literature is 23.8 kb, and this is a recurrent mutation in the French population (Mazoyer 2005, Rouleau 2007).

Deletion of the 5′ Region to Exon 2 (BRCA1)

By visual inspection, the mutation appeared as a deletion of the green signal S10B1, as well as a large genomic portion of the 5′ region upstream of BRCA1, including S11S1 and S12B1 (FIG. 4C). After measurement, the mutation was estimated to have a size of 37.1 kb, encompassing the portion of the BRCA1 gene that encodes exon 2, the entire NBR2 gene (signal S11B1), the genomic region between NBR2 and the pseudogene ψBRCA1 (signal S12B1), and a portion of ψBRCA1 (signal S13B1). Importantly, the reported size of this type of rearrangement is highly variable, originally in the range of 13.8 to 36.9 kb (Mazoyer 2005) and more recently between 40.4 and 58.1 kb (Rouleau 2007). Six different exon 1-2 deletions have been reported, 16 times, in a number of different populations (Sluiter 2010). The rearrangement reported here has been described three times with an identical size (36 934 bp). The hotspot for recombination is explained by the presence of ψBRCA1. Molecular combing proved capable of characterizing events even in this highly homologous region.

The results reported herein disclose and exemplify the development of a novel genetic test based on Molecular Combing for the detection of large rearrangements in the BRCA1 and BRCA2 genes. Large rearrangements represent 10-15% of deleterious germline mutations in the BRCA1 gene and 1-7% in the BRCA2 gene (Mazoyer, 2005). Specific high-resolution GMC were designed and were tested on a series of 16 biological samples; the robustness of the associated measurement strategy was statistically validated on 10 control samples, and 6 different large rearrangements were detected and characterized in samples from patients with a severe family history of breast cancer. The robustness of the newly designed GMC, devoid of repetitive sequences, is endorsed by the fact that our Molecular Combing method confirmed the results obtained with high-resolution zoom-in aCGH (11 k) on the same samples (Rouleau et al., 2007), with a resolution in the 1-2 kb range.

Tandem repeat duplications are the most difficult large rearrangements to detect. Contrary to other techniques, such as aCGH and MLPA, the capacity of Molecular Combing to visualize hybridized DNA probes at high resolution permits precise mapping and characterization of tandem repeat duplications, as shown here in case 1 (BRCA1 Dup Ex 13). aCGH can be used to determine the presence and size of duplications, but not the exact location and orientation of tandem repeat duplications. In PCR-based techniques such as MLPA, duplications are considered to be present when the ratio between the number of duplicated exons in the sample carrying a mutation and the number of exons in the control sample is at least 1.5, reflecting the presence of 3 copies of a specific exon in the mutated sample and 2 copies in the wild-type sample. The ratio of 1.5 is difficult to demonstrate unambiguously by MLPA, which often gives false-positive signals, as observed in case 1 (BRCA1 Dup Ex 13). The limits of MLPA have been underlined in several recent studies (Cavalieri et al., 2008), (Staaf et al., 2008). MLPA is limited to coding sequences and can also give false-negative scores, due to the restricted coverage of the 21 probes (Cavalieri et al., 2008). In addition, MLPA provides only limited information on the location of deletion or duplication breakpoints in the usually very large intronic or affected flanking regions, thus necessitating laborious mapping for sequence characterization of the rearrangements. Staaf et al recently suggested that MLPA should be regarded as a screening tool that needs to be complemented by other means of mutation characterization, such as a CGH (Staaf et al., 2008). We propose Molecular Combing as such a replacement technology for MLPA or aCGH, as it unambiguously identifies and visualizes duplications.

Another advantage of Molecular Combing as disclosed herein was its capacity to cover non-coding regions, including the 5′ region of the BRCA1 gene and the genomic region upstream of BRCA1 that comprises the NBR2 gene, the ψBRCA1 pseudogene and the NBR1 gene. Recent studies show that it is very difficult to design exploitable PCR or aCGH probes in this rearrangement-prone genomic region (Rouleau et al., 2007), (Staaf et al., 2008), because of the presence of duplicated regions and the high density of Alu repeats. Genomic rearrangements typically arise from unequal homologous recombination between short interspersed nuclear elements (SINEs), including Alu repeats, long interspersed nuclear elements (LINEs), or simple repeat sequences.

Molecular Combing permits precise physical mapping within this difficult regions, as shown here in cases three and two (BRCA1 Del Ex 2), where we measured mutation sizes of 38.5 kb and 37.1 kb, respectively. As cases 3 and 2 belong to the same family, the detected mutation was the same in both cases, as confirmed by aCGH (Rouleau et al., 2007). The measurement difference of 1.4 kb between these two cases is acceptable, being within the 1-2 kb definition range of the molecular combing assay. The mutation was originally described by Puget et al, who determined the mutation size (37 kb) with a first-generation molecular combing “color bar coding” screening method (Puget et al., 2002). Size estimated with aCGH was in the 40.4-58.1 kb range, because of the low density of exploitable oligonucleotide sequences in this genomic region and the reduced sensitivity of 22 some oligonucleotides due to sequence homology (Rouleau et al., 2007). Molecular combing can therefore be used for the analysis of hard-to-sequence genomic regions that contain large numbers of repetitive elements. Here we demonstrate that the high concentration of Alu sequences in BRCA1 does not represent an obstacle for molecular combing.

Detection of Previously Uncharacterized BRCA1 Large Rearrangements in Breast Cancer Patients

Further samples were tested, and we characterized by Molecular Combing rearrangements which other techniques had failed to accurately describe. One such example is detailed below.

Triplication of Exons 1a, 1b and 2 of BRCA1 and a Portion of NBR2.

We analyzed sample #7 (provided by the Institut Claudius Régaud, Toulouse, France) by Molecular Combing, using the set of probes described in FIG. 5. By visual inspection, two alleles of the BRCA1 gene were identified, differing in the length of the motif g7b1 which extends from the end of the S9B1 probe to the opposite end of the S11B1 probe. The mutation appears to be a triplication involving portions of the SYNT1 probe (SEQ ID 133) and the S10B1 probe, as was confirmed in probe color swapping experiments. This triplication of a DNA segment with a size comprised between 5 and 10 kb involves exons 1a, 1b and 2 of the BRCA1 gene and possibly part of the 5′ extremity of the NBR2 gene.

Such a triplication has not been reported in this genomic region yet. This may be due to the previous lack of relevant technologies to detect the mutation. Therefore, we designed tests specific to this mutation. These tests may be used to screen for this triplication or to confirm this triplication in samples where a rearrangement is suspected in this region. There are several types of possible tests, such as PCR, quantitative PCR (qPCR), MLPA, aCGH, sequencing . . . .

Results of quantification techniques, which provide a number of copies of a given sequence (qPCR, MLPA, aCGH, . . . ) will not provide direct assessment of the tandem nature of the additional copies of the sequence. The triplication reported here may be suspected when sequences within exons 1a, 1b and/or 2 of BRCA1 and/or the sequences between these exons are present in multiple (more than two per diploid genome) copies. Generally speaking, when these results are above the threshold determined for duplicated sequence (which have three copies in total of the duplicated sequence), the sample should be suspected to bear a triplication on a single allele (rather than duplications of the sequence in two separate alleles. Confirmation of the triplication and its tandem nature may be obtained either through a PCR test or through a Molecular Combing test as described in this and the examples section.

As this is a more direct method, we detail some PCR designs here, in the example sections. The man skilled in the art may adapt these tests through common, generally known, molecular biology methods, e.g. by modifying primer locations within the sequence ranges mentioned, and/or modifying experimental conditions (annealing temperature, elongation time, . . . for PCR). Also, these tests may be included in “multiplex” tests where other mutations are also sought. For example, one or several pair(s) of primers designed to detect the triplication and described below may be used simultaneously with one or several other pair(s) of primers targeting distinct amplicons. In addition to these adaptations, several common variants exist for the molecular tests described. Nevertheless, these variants remain functionally identical to the described tests and the adaptation of our designs to these variants is easily achievable by the man skilled in the art. For example, sequencing may be replaced by targeted resequencing, where the region of interest is isolated for other genomic regions before the sequencing step, so as to increase coverage in the region of interest. As another example, semi-quantitative PCR, where DNA is quantity after amplification is assessed by common agarose electrophoresis, may replace QMPSF.

These results demonstrate that the developed Molecular Combing platform is a valuable tool for genetic screening of tandem repeat duplications, CNVs, and other complex rearrangements in BRCA1 and BRCA2, such as translocations and inversions, particularly in high-risk breast cancer families.

A prominent application of the developed molecular diagnostic tool is as a predictive genetic test. However, the methods and tools disclosed herein may be applied as or in a companion diagnostic test, for instance, for the screening of BRCA-mutated cells in the context of the development of PARP inhibitors. Such a genetic test can be applied not only to clinical blood samples, but also to circulating cells and heterogeneous cell populations, such as tumor tissues.

EXAMPLES Example 1 Materials and Methods

Preliminary Patient Screening

The Genomic Morse Code was validated on 10 samples from patients with no deleterious mutations detected in BRCA1 or BRCA2 (control patients). The genetic test was validated on 6 samples from patients with positive family history of breast cancer and known to bear large rearrangements affecting either BRCA1 or BRCA2. Total human genomic DNA was obtained from EBV-immortalized lymphoblastoid cell lines. Preliminary screening for large rearrangements was performed with the QMPSF assay (Quantitative Multiplex PCR of Short Fluorescent Fragments) in the conditions described by Casilli et al and Tournier et al (Casilli et al., 2002) or by means of MLPA (Multiplex Ligation-Dependent Probe Amplification) using the SALSA MLPA kits P002 (MRC Holland, Amsterdam, The Netherlands) for BRCA1 and P045 (MRC-Holland) for BRCA2. All 16 patients gave their written consent for BRCA1 and BRCA2 analysis.

Molecular Combing

Sample Preparation

Total human genomic DNA was obtained from EBV-immortalized lymphoblastoid cell lines. A 45-μL suspension of 10⁶ cells in PBS was mixed with an equal volume of 1.2% Nusieve GTG agarose (Lonza, Basel, Switzerland) prepared in 1×PBS, previously equilibrated at 50° C. The plugs were left to solidify for 30 min at 4° C., then cell membranes are solubilised and proteins digested by an overnight incubation at 50° C. in 250 μL of 0.5 M EDTA pH 8.0, 1% Sarkosyl (Sigma-Aldrich, Saint Louis, Mo., USA) and 2 mg/mL proteinase K (Eurobio, Les Ulis, France), and the plugs were washed three times at room temperature in 10 mM Tris, 1 mM EDTA pH 8.0. The plugs were then either stored at 4° C. in 0.5 M7EDTA pH 8.0 or used immediately. Stored plugs were washed three times for 30 minutes in 10 mM Tris, 1 mM EDTA pH 8.0 prior to use.

Probe Preparation

All BRCA1 and BRCA2 probes were cloned into pCR2.1-Topo or pCR-XL-Topo (Invitrogen) plasmids by TOPO cloning, using PCR amplicons as inserts. Amplicons were obtained using bacterial artificial chromosomes (BACs) as template DNA. The following BACs were used: for BRCA1, the 207-kb BACRP11-831F13 (ch17: 41172482-41379594, InVitrogen, USA); and for BRCA2, the 172-kb BAC RP11-486017 (ch13: 32858070-33030569, InVitrogen, USA). See Tables 1 and 2 for primer sequences and probe coordinates. Primer sequences are referenced as SEQ ID 1 to SEQ ID 130. In some cases (as detailed in table 1), additional artificial sequences were added to the 5′ end of the primer for ease of cloning. These artificial sequences are SEQ ID 134 (ForwardPrimerPrefix) for forward primers and SEQ ID 135 (ReversePrimerPrefix) for forward primers, both containing a poly-A and a restriction site for, respectively, AscI and PacI.

SEQ ID 131 (BRCA1-1A), SEQ ID 132 (BRCA1-1B) and SEQ ID 133 (BRCA1-SYNT1) are examples of probe sequences.

Whole plasmids were used as templates for probe labeling by random priming. Briefly, for biotin (Biota) labeling, 200 ng of template was labeled with the DNA Bioprime kit (Invitrogen) following the manufacturers instructions, in an overnight labeling reaction. For Alexa-488 (A488) or digoxigenin (Dig) labeling, the same kit and protocol were used, but the dNTP mixture was modified to include the relevant labeled dNTP, namely Dig-11-dUTP (Roche Diagnostics, Meylan, France) or A488-7-OBEA dCTP (Invitrogen) and its unlabelled equivalent, both at 100 μM, and all other dNTPs at 200 μM. Labeled probes were stored at −20° C. For each coverslip, 5 ut of each labeled probe ( 1/10th of a labeling reaction product) was mixed with 10 μg of human Cot-1 and 10 μg of herring sperm DNA (both from Invitrogen) and precipitated in ethanol. The pellet was then resuspended in 22 μL of 50% formamide, 30% Blocking Aid (Invitrogen), 1×SSC, 2.5% Sarkosyl, 0.25% SDS, and 5 mM NaCl.

Genomic DNA Combing and Probe Hybridization

Genomic DNA was stained by 1 h incubation in 40 mM Tris, 2 mM EDTA containing 3 μM Yoyo-1 (Invitrogen, Carlsbad, Calif., USA) in the dark at room temperature. The plug was then transferred to 1 mL of 0.5 M MES pH 5.5, incubated at 68° C. for 20 min to melt the agarose, and then incubated at 42° C. overnight with 1.5 U beta agarase I (New England Biolabs, Ipswich, Mass., USA). The solution was transferred to a combing vessel already containing 1 ml of 0.5 M MES pH 5.5, and DNA combing was performed with the Molecular Combing System on dedicated coverslips (Combicoverslips) (both from Genomic Vision, Paris, France).

Combicoverslips with combed DNA are then baked for 4 h at 60° C. The coverslips were either stored at −20° C. or used immediately for hybridisation. The quality of combing (linearity and density of DNA molecules) was estimated under an epi-fluorescence microscope equipped with an FITC filter set and a 40× air objective. A freshly combed coverslip is mounted in 20 μL of a 1 ml ProLong-gold solution containing 1 μL of Yoyo-1 solution (both from Invitrogen). Prior to hybridisation, the coverslips were dehydrated by successive 3 minutes incubations in 70%, 90% and 100% ethanol baths and then air-dried for 10 min at room temperature. The probe mix (20 μL; see Probe Preparation) was spread on the coverslip, and then left to denature for 5 min at 90° C. and to hybridise overnight at 37° C. in a hybridizer (Dako). The coverslip was washed three times for 5 min in 50% formamide, 1×SSC, then 3×3 min in 2×SSC.

Detection was performed with two or three successive layers of flurophore or streptavidin-conjugated antibodies, depending on the modified nucleotide employed in the random priming reaction (see above). For the detection of biotin labeled probes the antibodies used were Streptavidin-A594 (InVitrogen, Molecular Probes) for the 1st and 3rd layer, biotinylated goat anti-Streptavidin (Vector Laboratories) for the 2nd layer; For the detection of A488-labelled probes the antibodies used were rabbit anti-A488 (InVitrogen, Molecular Probes) for the 1st and goat anti-rabbit A488 (InVitrogen, Molecular Probes) for the 2nd layer; For the detection of digoxygenin labeled probes the antibodies used were mouse anti-Dig (Jackson Immunoresearch) for the 1st layer, ratanti-mouse AMCA (Jackson Immunoresearch) for the 2nd layer and goat anti-mouse A350 (InVitrogen, Molecular Probes) for the 3rd Layer.

A 20 minute incubation step was performed at 37° C. in a humid chamber for each layer, and three successive 3 minutes washes in 2×SSC, 0.1% Tween at room temperature between layers. Three additional 3 minutes washes in PBS and dehydration by successive 3 minutes washes in 70%, 90% and 100% ethanol were performed before mounting the coverslip.

Image Acquisition

Image acquisition was performed with a customized automated fluorescence microscope (Image Xpress Micro, Molecular Devices, Sunnyvale, Calif., USA) at 40× magnification, and image analysis and signal measurement were performed with the software ImageJ (http://_rsbweb.nih.gov/ij) and JMeasure (Genomic Vision, Paris, France). Hybridisation signals corresponding to the BRCA1 and BRCA2 probes were selected by an operator on the basis of specific patterns made by the succession of probes. For all motifs signals belonging to the same DNA fibre, the operator set the ends of the segment and determined its identity and length (kb), on a 1:1 scale image. The data were then output as a spreadsheet. In the final analysis, only intact motif signals were considered, confirming that no fibre breakage had occurred within the BRCA1 or BRCA2 motifs.

Statistical Analysis

Molecular Combing allows DNA molecules to be stretched uniformly with a physical distance to contour length correlation of 1 μm, equivalent to 2 kb (Michalet et al., 1997). As a consequence, in the absence of large rearrangements, the derived stretching factor (SF) has a value close to 2 kb/μm (±0.2).

All 7 BRCA1 motifs (g1b1-g7b1) and all 5 BRCA2 motifs (g1b2-g5b2) were measured in all 20 biological samples. The mean value size of all motifs measured in the 10 healthy controls, including the associated statistical analysis, is reported in Table S1. The size of all motifs measured in the 6 breast cancer patients, including the associated statistical analysis, is reported in Table S2. For each motif, the following values were determined: the number of measured images (n), the theoretical calculated length (calculated (kb)), the mean measured length (p (kb)), the standard deviation (SD (kb)), the coefficient of variation (CV (%)), the difference between μ and calculated (delta), and the stretching factor (SF=(calculated/μ)×2) (Michalet et al., 1997). In the absence of mutations, delta values are comprised between −1.9 kb and 1.9 kb, and SF values are comprised between 1.8 and 2.2. The presence of a large rearrangement on BRCA1 or BRCA2 was first identified by visual inspection of the corresponding GMC. From numerous datasets, we established that in the presence of large rearrangements in both BRCA1 and BRCA2, delta≧2 kb (for duplications) or delta≦−2 kb (for deletions), and the corresponding SF≧2.3 kb/μm (for deletions) or SF≦1.7 kb/μm (for duplications). To confirm the presence of a large rearrangement, the motif (−s) of interest was (were) first measured on a total population of images (typically between 20 and 40), comprising wild-type (wt) and mutated (mt) alleles. In presence of large rearrangements, and aiming to measure the mutation size, the images were then divided in two groups, corresponding to the wt and the mt alleles. Within each of the two groups of n images, following values were calculated: μ (kb), SD (kb), CV (%). The μ value of the wild-type allele was then compared with the μ value of the mutated allele. To this aim, we calculated the standard error of the mean (SEM=SD/√n) and the 95% confidence interval (95% CI=μ+2×SEM). The mutation size was then calculated as a difference between the mean size of the two alleles: mutation size=μ(BRCA1^(mt))−μ(BRCA1^(wt)). The related error was calculated according to following formula:

error=(((μ^(mt)+2×SEM^(mt))−(μ^(wt)−2×SEM^(wt)))−((μ^(mt)−2×SEM^(mt))−(μ^(wt)+2×SEM^(wt))))/2.

Example 2 Comparison of Genetic Morse Code and Molecular Combing of the Invention to Prior Color Bar Code Procedure

Part 1. Previous Application of Molecular Combing on Characterization of BRCA1 and BRCA2 Large Rearrangements: Design of Low Resolution Color Bar Codes (CBCs)

Molecular Combing has already been used by Gad et al. (Gad GenChrCan 2001, Gad JMG 2002) to detect large rearrangements in the BRCA1 and BRCA2 genes. The hybridization DNA probes originally used were part of a low resolution “color bar coding” screening approach composed of cosmids, PACs and long-range PCR products. Some probes were small and ranged from 6 to 10 kb, covering a small fraction the BRCA1 and BRCA2 loci. Other probes were very big (PAC 103014 measuring 120 kb for BRCA1 and BAC 486017 measuring 180 kb for BRCA2) and were covering the whole loci, including all the repetitive sequences. Thus, no bioinformatic analysis to identify potentially disturbing repetitive sequences has been even performed. More importantly, no repetitive sequence has been ever excluded from the design of the CBCs. This often resulted in incomplete characterizations of the screened mutations (see Part 3). As a consequence, detection of the probes often resulted in the superposition of individual colored signals (e.g., yellow/white spots resulting from superposition of different colored signals) and in strong background noise, undermining the quality of the images and preventing the development of a robust strategy to measure the signals length. In addition, no DNA probe was r isolated and cloned in an insert vector. The BRCA1 Color Bare Code (CBC) was composed of only 7 DNA probes ((Gad, et al, Genes Chromosomes and cancer 31:75-84 (2001))), whereas the BRCA2 CBC was composed of only 8 DNA probes (Gad, et al, J Med Genet (2002)). This low number of DNA probes did not allow high resolution physical mapping.

Importantly, such a low resolution screening approach did not allow the unambiguous visualization of complex mutations, such as tandem repeat duplications or triplications. In contrast, full characterization of tandem repeat duplications and triplications is possible with the high-resolution GMC (see Example 1). Moreover, the accurate physical mapping of all the mutated exons was often problematic, requiring additional laborious sequencing experiments. This often resulted in incomplete characterizations of the screened mutations (see Chapter 3).

Part 2. New Application of Molecular Combing on Characterization of BRCA1 and BRCA2 Large Rearrangements: Design of High Resolution Genomic Morse Codes (GMCs) and Development of a Genetic Test.

An important point of novelty for the present invention is the design and cloning of high-resolution Genomic Morse Codes (GMC) for both BRCA1 and BRCA2 genomic regions. The BRCA1 GMC is composed of 35 DNA probes (FIG. 1), whereas the BRCA2 GMC is composed of 27 DNA probes (FIG. 2).

Comparative FIG. 1: in-silico generated (top) and microscopy observed (bottom) high resolution BRCA1 GMC.

Comparative FIG. 2: in-silico generated (top) and microscopy observed (bottom) high resolution GMC of BRCA2.

35 genomic regions in BRCA1 and 27 regions in BRCA2 devoid of repetitive sequences were identified, and were used to design and clone the corresponding DNA hybridization probes. All the details of the employed DNA hybridization probes (name, size, coordinates, color and the nature of the covered exons) are listed above. The cloned DNA probes allow the accurate physical mapping of deleted exons and permit the simultaneous detection of large rearrangements in BRCA1 and BRCA2. The above described improvement in resolution, permitted the inventors to translate their observations into the development of a robust predictive genetic test for breast and ovarian cancer (see example 1).

Part 3: High Resolution GMCs Allow the Unambiguous Detection and Visualization of Complex Mutation (e.g.: Tandem Repeat Duplications and Triplications) that can't be Characterized by Low Resolution CBCs

The following are selected examples of complex mutations that could not be characterized (or only partially) by low resolution CBC, but could be precisely and unambiguously characterized by high resolution GMC:

3.1 BRCA1 Dup Ex 18-20

CBC:

The image generated by Gad et al (case IC171712 in FIG. 1 of Gad et al, Oncogene 2001) has a low resolution and the nature and particularly the identity of the deleted exons cannot be defined by visual inspection. As a consequence, the size of the mutation has not been determined, confirming that the generated images were problematic for measurements.

GMC: (See Table S2 of Example 1)

By visual inspection, this mutation appears as a tandem duplication of the red signal S5B1. After measurement, the mutation was estimated to have a size of 6.7±1.2 kb, restricted to a portion of the genome that encodes for exons 18 to 20. The estimated mutation size is fully in line with the 8.7 kb reported in the literature (Staaf, 2008). Details on the measurement and statistical analysis can be found in Example 1.

Comparative FIG. 3: characterization of the BRCA1 mutation Dup ex 18-20 via CBC (top) and GMC (bottom).

3.2 BRCA1 Del Ex 8-13

CBC:

The image generated by Gad et al (case IC657 in FIG. 1 of Gad et al, Oncogene 2001) has a low resolution and the nature of the deleted exons cannot be unambiguously defined by visual inspection. The size of the mutation after measurement was 20.0±9.6 kb, having an important standard deviation.

GMC: (See FIG. 4B, Example 1)

By visual inspection, the mutation clearly appeared as a deletion of the blue signal S7B1, including a large genomic portion between signals S7B1 and S8B1. After measurement, the mutation was estimated to have a size of 20±2.8 kb, having a smaller error.

3.3 BRCA1 Dup Ex 13 (6.1 kb)

CBC:

No microscopy image related to mutation has been ever provided. The estimated mutation size was 5.8±1.8 kb (case IARC3653 in FIG. 3 of Gad et al, Oncogene 2001), but is not supported by visual inspection.

GMC: (see FIG. 4A, Example 1)

By visual inspection via Molecular Combing, this mutation appears as a partial tandem duplication of the blue signal S7B1. After measurement, the mutation was estimated to have a size of 6.1±1.6 kb, restricted to a portion of the DNA probe BRCA1-8 that encodes exon 13. The estimated mutation size is fully in line with the 6.1 kb reported in the literature (Puget, 1999), and according to the Breast Cancer Information Core database, this mutation belongs to the 10 most frequent mutations in BRCA1 (Szabo, 2000). Therefore, there is perfect correlation between the images and the measurements, and correlation with values present in literature. 3.4 Tandem repeat triplication of exons 1a, 1b and 2 of BRCA1 and a portion of NBR2.

CBC:

No tandem triplication has been ever reported using the CBC.

GMC:

By visual inspection via Molecular Combing, two alleles of the BRCA1 gene were identified in a sample provided by the Institut Claudius Regaud, Toulouse, France, differing in the length of the motif g7b1 which extends from the end of the S9B1 probe to the opposite end of the S11B1 probe. The mutation appeared to be a triplication involving portions of the SYNT1 and the S10B1 probe, as confirmed in probe color swapping experiments. This triplication of a DNA segment with a size comprised between 5 and 10 kb, and probably between 6 and 8 kb, involves exons 1a, 1b and 2 of the BRCA1 gene and possibly part of the 5′ extremity of the NBR2 gene.

The CBC would have at best detected this mutation as an increase of the length of a single probe, and thus would not have been able to characterize the mutation as a tandem triplication. Contrarily to Molecular Combing, none of the current molecular diagnostics technology, such as MLPA or aCGH, could assess whether the duplication or triplication is in tandem (within BRCA1) or dispersed (out of BRCA1). This observation makes a clear difference in terms of risk evaluation, since there is no evidence that repeated genomic portions out of the BRCA1 locus are clinically significant. Molecular Combing highlights that the mutation occurs within the BRCA1 gene, thus being of clinical significance.

The following important advantages of GMC compared to CBC are evident from the examples above:

-   -   high resolution visual inspection     -   precise mapping of mutated exons     -   precise measurement of mutation size with robust statistics     -   simultaneous detection of BRCA1 and BRCA2     -   detection of inversions and translocation     -   absence of disturbing repetitive sequence (Alu sequences) for         GMCs BRCA1 and BRCA2.

Tests Specific to Detect a Triplication in the 5′ Region of BRCA1

PCR tests to detect unambiguously the triplication described above or a close triplication may distinguish non triplicated from triplicated alleles through either one of two ways:

-   -   a—appearance of PCR fragments with the triplicated allele that         do not appear with a non-triplicated allele or;     -   b—change of size of a PCR fragment.

The organization of the sequences in a triplication may be used to design primer pairs such that the PCR amplification is only possible in a tandem repeat. If one of the primers is located in the amplified sequence and is in the same orientation as the BRCA gene (5′ to 3′) and the other is the reverse complementary of a sequence within the amplified sequence located upstream of the first primer (i.e. the direction from the location of the first to the second primer is the same as the direction from the 3′ to the 5′ end of the BRCA gene), the PCR in a non-mutated sample will not be possible as the orientation of the primers do not allow it. Conversely, in a triplicated sample, the first primer hybridizing on a repeat unit is oriented correctly relative to the second primer hybridizing in the repeat unit immediately downstream of the first primer's repeat unit. Thus, the PCR is possible. In a triplicated sample, two PCR fragments should be obtained using a pair of primers designed this way. In a sample with a duplication, only one fragment would appear. The size of the smaller PCR fragment (or the only fragment in the case of a duplication), s, is the sum of the following distances:

-   -   D, measured from the first (downstream) primer to the downstream         (3′ direction relative to the BRCA1 gene) breakpoint, and     -   U, measured from the second (upstream) primer to the upstream         (5′ direction relative to the BRCA1 gene) breakpoint.

This measurement thus provides a location range for both breakpoints, the downstream breakpoint being at a distance smaller than or equal to s from the location of the downstream primer (in the downstream direction) and the upstream breakpoint at a distance smaller than or equal to s from the location of the upstream primer (in the upstream direction). Besides, since the size of the triplicated sequence (L) is the sum of U+D and the distance between the two primers, L may be readily deduced from the size of the PCR fragment.

The size of the larger fragment is the sum of L and the size of the smaller fragment. Thus, by substracting the size of the smaller fragment from the size of the larger one, the size of the triplicated sequence is readily assessable in a second, independent assessment. This reduces the uncertainty on the location of the breakpoints. Thus, a test designed this way will allow a precise characterization of the triplication. Given the location of the triplication identified here, primer pairs used to detect the triplication could include combinations of one or several of the following downstream and upstream primers (the primer designed as the downstream primer is in the direct orientation relative to the BRCA1 gene and while the upstream primer is reverse complementary to the first strand of the BRCA1 gene). In choosing a combination of primers, in addition to the prescriptions below, one must choose the primer locations so the downstream primer is located downstream of the upstream primer:

A downstream primer may be located:

-   -   i) in the region between exons 2 and 3 of BRCA1, preferably at a         distance from 2-4 kb from the 3′ end of exon 2, more preferably         at a distance from 2.5-3 kb from the 3′ end of exon 2     -   ii) in the region between exons 2 and 3 of BRCA1, within 2 kb         from the 3′ end of exon 2, preferably within 1.5 kb and more         preferably within 1 kb from the 3′ end of exon 2

An upstream primer may be located:

-   -   i) in the region between the BRCA1 gene and the NBR2 gene,         within 2 kb from exon 1a of BRCA1, preferably within 1.5 kb and         more preferably within 1 kb of exon 1a of BRCA1;     -   ii) within exon 1a of BRCA1 or within exon 1b or in the region         between exons 1a and 1b;     -   iii) in the region between exons 1b and 2, or in exon 2, or in         the region between exons 2 and 3.

An example of such a combination is the primer pair consisting of primers BRCA1-Synt1-R (SEQ ID 126) and BRCA1-A3A-F (SEQ ID 25);

The combinations above are not meant to be exhaustive and the man skilled in the art may well choose other location for the upstream and downstream primers, provided the orientation and relative location of the primers is chosen as described. Several combinations of primers may be used in separate experiments or in a single experiment (in which case all of the “upstream” primers must be located upstream of all of the “downstream” primers. If more than three primers are used simultaneously (multiplex PCR°, the number of PCR fragments obtained will vary depending on the exact location of the breakpoint (no PCR fragment at all will appear in non mutated samples) and the characterization of the mutation will be difficult. Therefore, it is advisable to perform additional experiments with separate primer pairs if at least one fragment is observed in the multiplex PCR.

Importantly, with the design described in the preceeding paragraphs, the orientation of the triplicated sequence is of minor importance: indeed, in a triplication, at least two of the repeat units will share the same orientation and at least one PCR fragments should be amplified. This holds true for a duplication, as in the case of an inverted repeat, a PCR fragment would be obtained from a one of the primers hybridizing in two separate locations with reverse (facing) orientations, while a direct tandem repeat would generate a PCR fragment from the two primers as described above.

Another type of PCR test to reveal the triplication and its tandem nature requires the amplification of a fraction of or of the entire repeat array, using primer pairs spanning the repeated sequence (both primers remaining outside the amplified sequence), or spanning a breakpoint (one primer is within and the other outside the amplified sequence) or entirely included in the amplified sequence. These tests will generate a PCR fragment of given size in a normal sample, while in a sample with a triplication on one allele, one or more additional PCR fragment will appear, including one the size of the “normal” fragment plus twice the size of the repeat sequence. If a mutation is present, these tests will often lead to results than can have several interpretations. If a single experiment is performed and reveals a mutation, a (series of) complementary test(s) may be performed following the designs presented herein to confirm the correct interpretation. Given the location of the triplication identified here, primer pairs used to detect the triplication could include a combination of one or several of the following primers, with at least one down stream and one upstream primer. The primer designed as the downstream primer is reverse complementary relative to the BRCA1 gene sequence and while the upstream primer is in direct orientation relative to the BRCA1 gene. In choosing a combination of primers, in addition to the prescriptions below, one must choose the primer locations so the downstream primer is located downstream of the upstream primer:

A downstream primer may be located:

-   -   i) in exon 3 of the BRCA1 gene; or     -   ii) in the region between exons 2 and 3 of BRCA1, preferably         more than 2 kb and less than 10 kb from the 3′ end of exon 2,         more preferably more than 3 kb and less than 8 kb and even more         preferably more than 4 kb and less than 6 kb from the 3′ end of         exon 2.

An upstream primer may be located:

-   -   i) in the region between the BRCA1 gene and the NBR2 gene, less         than 10 kb from exon 1a of BRCA1 and more than 1 kb from exon 1a         of BRCA1, preferably more less than 8 kb than 2 kb and more         preferably less than 6 and more than 4 kb of exon 1a of BRCA1;         or     -   ii) in exon 1a, exon 1b or in the region between exons 1a and 1b         of BRCA1; or     -   iii) in exon 2 or in the region between exons 1b and 2 of BRCA1         or in the region between exons 2 and 3.     -   iii)     -   iv)     -   Examples of such combinations are the primer pairs consisting of         primers BRCA1-A3A-F (SEQ ID 25) and BRCA1-A3A-R (SEQ ID 26) and         of primers BRCA1-Synt1-F (SEQ ID 125) and BRCA1-Synt1-R (SEQ ID         126)     -   v) a downstream primer as described in i) and an upstream primer         as described in ii)     -   vi) a dowstream primer as described in i) and an upstream primer         as described in iii)     -   vii) a dowstream primer as described in ii) and an upstream         primer as described in i)

Specific Embodiments of the Invention Include the Following:

1. A nucleic acid composition for detecting simultaneously one or more large or complex mutations or genetic rearrangements in the locus BRCA1 or BRCA2 comprising at least two colored-labeled probes containing more than 200 nucleotides and specific of each said gene, said probes being visually detectable at high resolution and free of repetitive nucleotidic sequences.

2. A nucleic acid composition according to embodiment 1 for detecting simultaneously one or more large or complex mutations or genetic rearrangements in the locus BRCA1 or BRCA2 comprising at least three colored-labeled probes containing more than 200 nucleotides and specific of each said gene, said probes being visually detectable at high resolution and free of repetitive nucleotidic sequences.

3. A nucleic acid composition according to embodiments 1 or 2 for detecting simultaneously one or more large or complex mutations or genetic rearrangements in BRCA1 or BRCA2 gene comprising at least three color-labeled probes containing more than 600 nucleotides and specific of each said gene, said probes being visually detectable at high resolution and free of repetitive nucleotidic sequences.

4. A composition according embodiments 1, 2 or 3, wherein the probes are all together visualized on a monostranded-DNA fiber or on a polynucleotidic sequence of interest or on a genome to be tested.

5. A composition according embodiments 1, 2, 3 or 4 comprising at least five color-labeled signal probes specific of BRCA1 or BRCA2 locus allowing detection of the following mutations: duplication, deletion, inversion, insertion, translocation or large rearrangement.

6. A composition according embodiments 1 to 4 comprising at least seven color-labeled signal probes specific of BRCA1 or BRCA2 locus allowing to detect following mutations: duplication, deletion, inversion, insertion, translocation or large rearrangement.

7. A composition according embodiments 1 to 4 comprising at least nine color-labeled signal probes specific of BRCA1 or BRCA2 locus allowing to detect following mutations: duplication, triplication, deletion, inversion, insertion, translocation or large rearrangement.

8. A composition according embodiments 1 to 7 comprising at least fourteen color-labeled signal probes specific of BRCA1 or BRCA2 locus allowing to detect following mutations: duplication, triplication, deletion, inversion, insertion, translocation or large rearrangement.

9. A composition according embodiments 1 to 8 comprising at least eighteen color-labeled signal probes specific of BRCA1 or BRCA2 locus allowing to detect following mutations: duplication, triplication, deletion, inversion, insertion, translocation or large rearrangement.

10. A composition according to embodiments 1 to 9 wherein the genetic rearrangement or mutation detected is more than 1.5 kilobase (kb).

11. A predictive genetic test of susceptibility of breast or ovarian cancer in a subject involving the detection (presence or absence) and optionally the characterization of one or more specific large genetic rearrangement or mutation in the coding or non coding sequences of the BRCA1 or BRCA2 locus, the rearrangement being visualized by any of the composition according to embodiments 1 to 10.

12. A method of detection for the sensitivity of a subject to a therapeutic procedure comprising the identification of one or more genetic rearrangements or mutations in the coding or non-coding sequences of BRCA1 or BRCA2 gene or locus by visualizing by molecular combing said genetic rearrangement by using any of the composition according to embodiments 1 to 10.

13. A method of detection of at least one large genetic rearrangement or mutation by molecular combing technique in a fluid or circulating cells or a tissue of a biological sample comprising the steps of

a) contacting the genetic material to be tested with at least two colored labeled probes according to embodiments 1 to 10 visualizing with high resolution the hybridization of step a) and optionally

b) comparing the result of step b) to the result obtained with a standardized genetic material carrying no rearrangement or mutation in BRCA1 or BRCA2 gene or locus.

14. A composition comprising:

two or more oligonucleotide probes according to embodiments 1 to 10;

probes complementary to said oligonucleotide probes;

probes that hybridize to said probes of embodiments 1 to 10 under stringent conditions;

probes amplified by PCR using pairs of primers described in Tables 1 or 2 (SEQ ID 1 to SEQ ID 130); or

probes comprising BRCA1-1A (SEQ ID NO: 131), BRCA1-1B (SEQ ID NO: 132), or BRCA1-SYNT1 (SEQ ID NO:133)

15. A set of primers selected from the group of primers consisting of SEQ ID 1 to SEQ ID 70 and SEQ ID 125 to SEQ ID 130 for BRCA1

16. A set of primers selected from the group of primers consisting of SEQ ID 71 to SEQ ID 124 for BRCA2.

17. An isolated or purified probe produced by amplifying BRCA1 or BRCA2 coding, intron or flanking sequences using a primer pair of embodiment 15 or 16.

18. An isolated or purified probe comprising a polynucleotide sequence of SEQ ID NO: 131 (BRCA1-1A), SEQ ID NO: 132 (BRCA1-1B) or SEQ ID NO: 133 (SYNT1), or that hybridizes to SEQ ID NO: 131 or to SEQ ID NO: 132 or to SEQ ID NO: 133 under stringent conditions.

19. A composition comprising at least two polynucleotides each of which binds to a portion of the genome containing a BRCA1 and/or BRCA2 gene, wherein each of said at least two polynucleotides contains at least 200 contiguous nucleotides and contains less than 10% of Alu repetitive nucleotidic sequences.

20. The composition of embodiment 19, wherein said at least two polynucleotides bind to a portion of the genome containing BRCA1.

21. The composition of embodiment 19, wherein said at least two polynucleotides bind to a portion of the genome containing BRCA2.

22. The composition of embodiment 19, wherein each of said at least two polynucleotides contains at least 500 up to 6,000 contiguous nucleotides and contains less than 10% of Alu repetitive nucleotidic sequences.

23. The composition of embodiment 19, wherein the at least two polynucleotides are each tagged with a detectable label or marker.

24. The composition of embodiment 19, comprising at least two polynucleotides that are each tagged with a different detectable label or marker.

25. The composition of embodiment 19, comprising at least three polynucleotides that are each tagged with a different detectable label or marker.

26. The composition of embodiment 19, comprising at least four polynucleotides that are each tagged with a different detectable label or marker.

27. The composition of embodiment 19, comprising three to ten polynucleotides that are each independently tagged with the same or different visually detectable markers.

28. The composition of embodiment 19, comprising eleven to twenty polynucleotides that are each independently tagged with the same or different visually detectable markers.

29. The composition of embodiment 19, comprising at least two polynucleotides each tagged with one of at least two different detectable labels or markers.

30. A method for detecting a duplication, triplication, deletion, inversion, insertion, translocation or large rearrangement in a BRCA1 or BRCA2 locus, BRCA1 or BRCA gene, BRCA1 or BRCA flanking sequence or intron, comprising: isolating a DNA sample, molecularly combing said sample, contacting the molecularly combed DNA with the composition of embodiment 5 as a probe for a time and under conditions sufficient for hybridization to occur, visualizing the hybridization of the composition of embodiment 5 to the DNA sample, and comparing said visualization with that obtain from a control sample of a normal or standard BRCA1 or BRCA2 locus, BRCA1 or BRCA gene, BRCA1 or BRCA flanking sequence or intron that does not contain a rearrangement or mutation.

31. The method of embodiment 30, wherein said probe is selected to detect a rearrangement or mutation of more than 1.5 kb.

32. The method of embodiment 30, further comprising predicting or assessing a predisposition to ovarian or breast cancer based on the kind of genetic rearrangement or mutation detected in a coding or noncoding BRCA1 or BRCA 2 locus sequence.

33. The method of embodiment 30, further comprising determining the sensitivity of a subject to a therapeutic treatment based on the kind of genetic rearrangement or mutation detected in a coding or noncoding BRCA1 or BRCA 2 locus sequence.

34. A kit for detecting a duplication, deletion, triplication, inversion, insertion, translocation or large rearrangement in a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron comprising at least two polynucleotides each of which binds to a portion of the genome containing a BRCA1 or BRCA2 gene, wherein each of said at least two polynucleotides contains at least 200 contiguous nucleotides and is free of repetitive nucleotidic sequences, wherein said at least two or polynucleotides are tagged with visually detectable markers and are selected to identify a duplication, deletion, inversion, insertion, translocation or large rearrangement in a particular segment of a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron; and optionally a standard describing a hybridization profile for a subject not having a duplication, deletion, inversion, insertion, translocation or large rearrangement in a BRCA1 or BRCA2 locus, BRCA1 or BRCA gene, BRCA1 or BRCA flanking sequence or intron; one or more elements necessary to perform Molecular Combing, instructions for use, and/or one or more packaging materials.

35. The kit of embodiment 34, wherein said at least two or polynucleotides are selected to identify a duplication, deletion, inversion, insertion, translocation or large rearrangement in a particular segment of a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron associated with ovarian cancer or breast cancer.

36. The kit of embodiment 34, wherein said at least two or polynucleotides are selected to identify a duplication, deletion, inversion, insertion, translocation or large rearrangement in a particular segment of a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron associated with a kind of ovarian cancer or breast cancer sensitive to a particular therapeutic agent, drug or procedure.

37. A method for detecting an amplification of a genomic sequence spanning the 5′ end of the BRCA1 gene and consisting of at least three copies of the sequence in a sample containing genomic DNA. Accordingly, the invention relates in particular to a method for in vitro detecting in a sample containing genomic DNA, a repeat array of multiple tandem copies of a repeat unit consisting of genomic sequence spanning the 5′ end of the BRCA1 gene wherein said repeat array consists of at least three copies of the repeat unit and said method comprises:

-   -   providing conditions enabling hybridization of a first primer         with the 5′ end of the target genomic sequence and hybridization         of a second primer with the 3′ end of said target sequence, in         order to enable polymerization by PCR starting from said         primers;     -   amplifying the sequences hybridized with the primers;     -   detecting, in particular with a probe, the amplicons thereby         obtained and determining their size or their content, in         particular their nucleotide sequence.

38. A method of embodiment 37, where the amplified sequence is at least 2 kb long.

39. A method of embodiment 37, where the amplified sequence is at least 5 kb long.

40. A method of embodiment 37, where the amplified sequence is at most 20 kb long.

41. A method of embodiment 37, where the amplified sequence is at most 10 kb long.

42. A method of embodiment 37, where the amplified sequence is at least 2 kb and at most 20 kb long.

43. A method of embodiment 37, where the amplified sequence is at least 5 kb and at most 10 kb long.

44. A method of any one of embodiments 37 to 43 where the amplified sequence comprises at least one of exons 1a, 1b and 2 of the BRCA1 gene.

45. A method of any one of embodiments 37 to 43 where the amplified sequence comprises exons 1a, 1 b and 2 of the BRCA1 gene.

46. A method of any one of embodiments 37-45 where the detection of the gene amplification is achieved by quantifying copies of a sequence included in the amplified region.

47. A method of any one of embodiments 37-46 where the detection of the gene amplification is achieved by measuring the size of a genomic sequence encompassing the amplified sequence.

48. A method of any one of embodiments 37-47 where the detection of the gene amplification is achieved by making use of polymerase chain reaction or other DNA amplification techniques.

49. A method of any one of embodiments 37 to 48 where the detection of the gene amplification is achieved by quantitative polymerase chain reaction

50. A method of any one of embodiments 37-48 where the detection of the gene amplification is achieved by multiplex, ligation-dependent probe amplification (MLPA).

51. A method of any one of embodiments 37-48 where the detection of the gene amplification is achieved by array-based comparative genomic hybridization (aCGH).

52. A method of any one of embodiments 37-48 where the detection of the gene amplification is achieved by quick multiplex PCR of short fragments (QMPSF)

53. A method of any one of embodiments 37-48 wherein the downstream and upstream primers are respectively selected from the group of:

for a downstream primer:

-   -   a polynucleotide sequence in the region between exons 2 and 3 of         BRCA1, preferably at a distance from 2-4 kb from the 3′ end of         exon 2, more preferably at a distance from 2.5-3 kb from the 3′         end of exon 2 or     -   a polynucleotide sequence in the region between exons 2 and 3 of         BRCA1, within 2 kb from the 3′ end of exon 2, preferably within         1.5 kb and more preferably within 1 kb from the 3′ end of exon 2         for an upstream primer:     -   a polynucleotide sequence in the region between the BRCA1 gene         and the NBR2 gene, within 2 kb from exon 1a of BRCA1, preferably         within 1.5 kb and more preferably within 1 kb of exon 1a of         BRCA1 or,     -   a polynucleotide sequence within exon 1a of BRCA1 or within exon         1b or in the region between exons 1a and 1b or,     -   a polynucleotide sequence in the region between exons 1b and 2,         or in exon 2, or in the region between exons 2 and 3

54. A method of any one of embodiments 37-48 using two or more primers chosen from BRCA1-A3A-F (SEQ ID 25), BRCA1-A3A-R (SEQ ID 26), BRCA1-Synt1-F (SEQ ID 125) and BRCA1-Synt1-R (SEQ ID 126) or their reverse complementary sequences. 55. A method of any one of embodiments 37-48 using the Synt 1 probe (SEQ ID NO: 133).

TABLE 1 Description of the DNA probes encoding the BRCA1 GMC Probe Probe size Forward Reverse BRCA1 name (bp) Primer¹ Primer² Start³ End³ Signal Motif Color^(d) Gene Exons BRCA1-1A 3548 aaaaggcgcgccG aaaattaattaaG 4237 7784 S1B1 G GGACGGAAAGCTA GGCAGAGGTGACA TGATGT GGTCTA BRCA1-1B 3561 aaaaggcgcgccC aaaattaattaaA 7842 11402 S1B1 G CTCTGACCTGATC TCAGCAACAGTCC CCTTGA CATTCC BRCA1-2 1900 aaaaggcgcgccG aaaattaattaaG 12936 14935 S1B1 G CCCAGACTAGTGT GCATGAGGCAGCA TTCTTAACC ATTTAG BRCA1-3 4082 aaaaggcgcgccT aaaattaattaaG 20012 24093 S2B1 g1b1 R BRCA1 25 + 26 CTTTGAATCTGGG CTGTTGCTTTCTT CTCTGC TGAGGTG BRCA1-4 2600 aaaaggcgcgccC aaaattaattaaC 28528 31129 S3B1 g2b1 R BRCA1 22 + 23 ACAGGTATGTGGG CTCTGTTGATGGG CAGAGA GTCATAG BRCA1-5 1400 aaaaggcgcgccT aaaattaattaaC 38009 42947 S4B1 g3b1 G BRCA1 TTGGTAGACCAGG AAATTATGTGTGG TGAAATGA AGGCAGA BRCA1-6 2924 aaaaggcgcgccG aaaattaattaaA 45870 45898 S5B1 g3b1 G BRCA1 19 AAGAACGTGCTCT AAGTCTGATAACA TTTCACG GCTCCGAGA BRCA1-7 2200 aaaaggcgcgccT aaaattaattaaC 48151 50350 S6B1 g3b1 G BRCA1 15 + 16 + 17 TCGATTCCCTAAG ACAGTTCTGTGTA ATCGTTTC ATTTAATTTCGAT BRCA1-8 3839 aaaaggcgcgccA aaaattaattaaT 58754 62592 S7B1 g4b1 B BRCA1 13 + 14 GGGAAGGCTCAGA GCCATAGATAGAG TACAAAC GGCTTTTT BRCA1-9 2688 aaaaggcgcgccG aaaattaattaaT 64151 66836 S7B1 g4b1 B BRCA1 CCATCTTCTTTCT TGACCTATTGCTG CCTGCT AATGTTGG BRCA1-11 2917 aaaaggcgcgccT aaaattaattaaG 83652 86568 S8B1 g5b1 B BRCA1 5 + 6 + 7 TTTACCAAGGAAG CTTGATCACAGAT GATTTTCG GTATGTATGAGTT BRCA1-12 2014 aaaaggcgcgccC aaaattaattaaT 93876 95889 S9B1 g6b1 B BRCA1 3 CCCAGGGCTTTAA AGGGGTGGATATG AGGTTA GGTGAA BRCA1-13A 1279 aaaaggcgcgcca aaaattaattaag 103601 104879 S10B1 g7b1 G BRCA1 1a + 1b + 2 cttcttcaacgcg acaggctgtgggg aagagc tttct BRCA1-15 3563 aaaaggcgcgccT aaaattaattaaT 113539 117101 S11B1 R NBR2 ATCTGCTGGCCAC CTCGAGCCTTGAA TTACCA CATCCT BRCA1-16 965 aaaaggcgcgccC aaaattaattaaA 117852 118816 S11B1 R NBR2 GCTCAGCTTTCAT AACGTTCACATGT TCCAGT ATCCCCTAA BRCA1-17 1574 aaaaggcgcgccC aaaattaattaaC 119183 120756 S11B1 R NBR2 CTGGCCAGTACCC TGAGCCCAGAGTT AGTAGT TCTGCT BRCA1-18 1376 aaaaggcgcgccG aaaattaattaaG 127190 128565 S12B1 B GGCCCAAAAACCA GGATTGAGCGTTC GTAAGA ACAGAT BRCA1-19 1969 aaaaggcgcgccG aaaattaattaaT 130024 131891 S12B1 B CCATCCAGTCCAG GCAGTTCTACCCT TCTCAT CCACTTG BRCA1-22 3912 aaaaggcgcgccC aaaattaattaaG 148370 152281 S13B1 G ΨBRCA1 + GGGTAAGTGGTGA AACTGTCTTTAAA NBR1 GCTTTC GGCACTTTTT BRCA1-23 2990 aaaaggcgcgccT aaaattaattaaT 154738 157727 S14B1 R NBR1 GGCTAGTGTTTTG TCAGTGTTGCTTC GCCTGT TCCATTTC BRCA1-24 1813 aaaaggcgcgccT aaaattaattaaA 158538 160350 S14B1 R NBR1 GTCAGACTAGCCA AGCGCTTCTTCAT CAGTACCA ATTCTCC BRCA1-25 735 aaaaggcgcgccA aaaattaattaaG 165696 166430 S15B1 G NBR1 CCACACTCTTCTG GCACATGTACACC TTTTGATGT ATGGAA BRCA1-26 3233 aaaaggcgcgccT aaaattaattaaT 167936 171168 S15B1 G NBR1 TGTGTAGGTTGCC TCAGAGAGCTGGG CGTTC CCTAAA BRCA1-27 2419 aaaaggcgcgccg aaaattaattaag 172299 174717 S15B1 G NBR1 gaggcaatctgga gatccatgattgc attgaa tgcttt BRCA1-29 970 aaaaggcgcgccC aaaattaattaaT 277732 278701 S16B1 B CCTCTAGATACTT CTGGCAGTCACAA GTGTCCTTTTG TTCAGG BRCA1-30 951 aaaaggcgcgccT aaaattaattaaT 281267 282217 S16B1 B CCCATGACTGCAT TGAGATCAGGTCG CATCTT ATTCCTC BRCA1-31 629 aaaaggcgcgccA aaaattaattaaC 282779 283407 S16B1 B AAACTCAACCCAA CAAGAATCACGAA ACAGTCA GAGAGAGA BRCA1-32 601 aaaaggcgcgccG aaaattaattaaG 283805 284405 S16B1 B ACCTCATAGAGGT CTCAAAGCCTTTA AGTGGAAAGAA GAAGAAACA BRCA1-33 648 aaaaggcgcgccG aaaattaattaaC 284755 285402 S16B1 B CACTGGGGAAAAG TCTTCAACCCAGA GTAGAA CAGATGC BRCA1-34 962 aaaaggcgcgccC aaaattaattaaC 289229 290190 S17B1 B AATACCCAATACA TGGGGATACTGAA ATGTAAATGC ACTGTGC BRCA1-35 4638 aaaaggcgcgccA aaaattaattaaT 290944 295581 S17B1 TMEM TCAAGAAGCCTTC CCTTGGACGTAAG 106A CCAGGT GAGCTG BRCA1-36 2944 aaaaggcgcgccT aaaattaattaaG 296903 299846 S17B1 B TMEM TCAGAACTTCCAA ATGGAGCTGGGGT 106A ATACGGACT GAAAT BRCA1-37 1302 aaaaggcgcgccC aaaattaattaaC 302021 303322 S18B1 G GTGAGATTGCTCA AAGGCATTGGAAA CAGGAC GGTGTC BRCA1-38 1464 aaaaggcgcgccA aaaattaattaaT 304919 306382 S18B1 G GAGGAATAGACCA CCTCCAGCACTAA TCCAGAAGT AAACTGC Notes: ¹12 bases (aaaaggcgcgcc) containing the restriction site sequence for AscI (GGCGCGCC) have been added for cloning purposes ²12 bases (aaaattaattaa) containing the restriction site sequence for PacI (TTAATTAA) have been added for cloning purposes ³cordinates relative to BAC RP11-831F13, according to NCBI Build 36.1 (hg18); ⁴B = blue, G = green, R = red

TABLE 2 Description of the DNA probes encoding the BRCA2 GMC Probe Probe size Forward Reverse BRCA2 name (bp) primer primer Start¹ End¹ Signal Motif Color² Gene Exons BRCA2-1 2450 AAATGGAGGTCAG TGGAAAGTTTGG 39 2488 S1B2 R GGAACAA GTATGCAG BRCA2-2 4061 TCTCAATGTGCAA TCTTGACCATGT 3386 7446 S1B2 R GGCAATC GGCAAATAA BRCA2-3a 3822 AATCACCCCAACC GCCCAGGACAAA 8935 12756 S1B2 R TTCAGC CATTTTCA BRCA2-3b 3930 CCCTCGCATGTAT CTCCTGAAGTCC 12808 16737 S1B2 R GATCTGA TGGAAACG BRCA2-3c 3953 TGAAATCTTTTCC AGATTGGGCACA 16756 20708 S1B2 R CTCTCATCC TCGAAAAG BRCA2-5 1903 GGTCTTGAACACC CACTCCGGGGGT 31031 32933 S2B2 g1b2 B BRCA2 1 + 2 TGCTACCC CCTAGAT BRCA2-6 4103 TCTTTAACTGTTC TGGCTAGAATTC 35073 39175 S2B2 g1b2 B BRCA2  3 TGGGTCACAA AAAACACTGA BRCA2-7 1854 TTGAAGTGGGGTT CCAGCCAATTCA 39617 41470 S2B2 g1b2 B BRCA2  4 TTTAAGTTACAC ACATCACA BRCA2-11 5206 TTGGGACAATTCT TGCAGGTTTTGT 52411 57616 S3B2 g2b2 G BRCA2 11 GAGGAAAT TAAGAGTTTCA BRCA2-12 5734 TGGCAAATGACTG TCTTGAAGGCAA 59208 64941 S4B2 g2b2 G BRCA2 12 + 13 CATTAGG ACTCTTCCA BRCA2-13 3251 GGAATTGTTGAAG ACCACCAAAGGG 68200 71450 S5B2 g3b2 R BRCA2 14 TCACTGAGTTGT GGAAAAC BRCA2-14 1681 CAAGTCTTCAGAA TAAACCCCAGGA 72505 74185 S5B2 g3b2 R BRCA2  15 + 16* TGCCAGAGA CAAACAGC BRCA2-15 4216 GGCTGTTTGTTGA GAAACCAGGAAA 76757 80972 S6B2 g3b2 R BRCA2 17 + 18 GGAGAGG TGGGGTTT BRCA2-18 2572 TGTTAGGGAGGAA GGATGTAACTTG 93846 96417 S7B2 g4b2 R BRCA2 22 + 23 + GGAGCAA TTACCCTTGAAA 24 BRCA2-19 2125 TCAATAGCATGAA GAGGTCTGCCAC 96951 99075 S7B2 g4b2 R BRCA2 TCTGTTGTGAA AAGTTTCC BRCA2-20 2559 GGCCCACTGGAGG TTCCTTTCAATT 99537 102095 S7B2 g4b2 R BRCA2  25* TTTAAT TGTACAGAAACC BRCA2-21 1568 TGAATCAATGTGT GTGTAGGGTCCA 102609 104176 S8B2 g5b2 B BRCA2 GTGTGCAT GCCCTATG BRCA2-22a 3787 CTGAGGCTAGGAA CTGAGGCTAGGA 104612 108398 S8B2 g5b2 B BRCA2 AGCTGGA AAGCTGGA BRCA2-22b 3606 GGTTTATCCCAGG AGAAAATGTGGG 108408 112013 S8B2 g5b2 B BRCA2 26 ATAGAATGG GTGTAAACAG BRCA2-25 5052 CAGCAAACTTCAG GGGACATGGCAA 123134 128185 S9B2 R CCATTGA CCAAATAC BRCA2-26 2353 GCACTTTCACGTC CGTCGTATTCAG 130493 132845 S10B2 R CTTTGGT GAGCCATT BRCA2-27 2058 CCCAGCTGGCAAA TCGGAGGTAATT 133176 135233 S10B2 R CTTTTT CCCATGAC BRCA2-28a 4158 TCAAGAGCCATGC AGGTAGGGTGGG 137121 141278 S11B2 R TGACATC GAAGAAGA BRCA2-29 2335 TGAGTCTACTTTG TTTTGCTTTCGG 153394 155728 S12B2 G CCCATAGAGG GAGCTTTA BRCA2-30 2121 TTTTTGCCTGCTT GGTTTTTAAACC 160291 161435 S13B2 B CATCCTC TGCACATGAA BRCA2-31 4803 TGAAATTTTGTTA TTTGAAATCTGT 161435 166237 S13B2 B TGTGGTGCAT GGAGGTCTAGC BRCA2-32 2609 GTACCAAGGGTGG ATGGTGTTGGTT 169818 172426 S14B2 G CAGAAAG GGGTAGGA Notes: ³cordinates relative to BAC RP11-486017, according to NCBI Build 36.1 (hg18) ⁴B = blue, G = green, R = red

TABLE 3 Total Alu sequences in probes  30 (10%) Total Alu sequences in excluded regions 270 (90%) position in repeat position in query sequence (hg18) matching repeat (left) end begin linkage Alu seq score % div. % del. % ins. begin end (left) + repeat class/family begin end (left) id (count) excluded region 1 2519 7.1 1.0 0.0 132 441 −308672 + AluSp SINE/Alu 1 313 0 1 7 25 72.0 0.0 0.0 1136 1160 −307953 + AT_rich Low_Cplxty 1 25 0 2 22 58.3 0.0 0.0 1627 1662 −307451 + GC_rich Low_Cplxty 1 36 0 3 223 19.3 3.5 0.0 1708 1764 −307349 + (CGG)n Simple 2 60 0 4 21 57.1 0.0 0.0 1959 1986 −307127 + GC_rich Low_Cplxty 1 28 0 5 2280 7.5 2.7 0.7 2142 2434 −306679 + AluSz SINE/Alu 1 299 −13 6 2216 10.4 0.0 1.4 2436 2733 −306380 + AluSx1 SINE/Alu 1 294 −18 7 2480 4.4 2.0 0.3 2734 3026 −306087 + AluY SINE/Alu 1 298 −13 8 1117 15.8 0.6 0.0 3305 3475 −305638 C AluJr SINE/Alu −11 301 130 9 364 13.5 0.0 0.0 3482 3533 −305580 C MER66A LTR/ERV1 −140 338 287 10 749 11.9 5.9 0.8 3557 3674 −305439 C AluJr SINE/Alu −187 125 2 9 1741 6.0 17.9 1.0 3746 3996 −305117 C AluY SINE/Alu −18 293 1 11 probe 1A 273 26.3 2.9 0.8 4677 4880 −304233 + G-rich Low_Cplxty 1 208 0 12 1 22 40.9 0.0 0.0 5327 5348 −303765 + GC_rich Low_Cplxty 1 22 0 13 2331 9.6 0.7 0.3 5904 6205 −302908 + AluSx SINE/Alu 1 303 −9 14 excluded region 2 0 probe 1B 2512 6.3 0.3 3.2 9150 9467 −299646 + AluY SINE/Alu 1 309 −2 15 2 313 24.8 17.9 0.0 9930 10046 −299067 C L2b LINE/L2 0 3375 3238 16 374 31.1 1.9 6.6 10058 10260 −298853 C L2b LINE/L2 −179 3208 3005 16 958 15.6 0.0 7.1 10508 10687 −298426 + FRAM SINE/Alu 8 175 −1 17 excluded region 3 1420 7.5 0.0 0.6 11598 11771 −297342 C AluSc SINE/Alu −2 307 135 18 7 2332 8.4 0.7 0.3 11783 12078 −297035 C AluSp SINE/Alu −16 297 1 19 486 10.1 0.0 15.1 12079 12129 −296984 C AluSc SINE/Alu −218 91 47 18 1515 13.5 0.9 0.5 12130 12344 −296769 C AluSx SINE/Alu −94 218 3 20 2169 8.4 1.4 1.7 12353 12507 −296606 C AluY SINE/Alu −20 291 133 21 2672 4.7 0.0 0.0 12508 12807 −296306 C AIuY SINE/Alu −11 300 1 22 2169 8.4 1.4 1.7 12808 12941 −296172 C AluY SINE/Alu −179 132 3 21 probe 2 2169 8.4 1.4 1.7 12808 12941 −296172 C AluY SINE/Alu −179 132 3 21 2 486 10.1 0.0 15.1 12942 12979 −296134 C AluSc SINE/Alu −177 132 99 18 381 34.8 4.9 0.6 13095 13256 −295857 + MIRc SINE/MIR 18 186 −82 23 219 29.5 2.8 2.8 13304 13411 −295702 C L2c LINE/L2 −202 3185 3078 24 449 3.2 0.0 0.0 13485 13546 −295567 + SVA _E Other 1318 1379 −3 25 601 28.4 18.6 0.0 14578 14771 −294342 + MIRb SINE/MIR 24 253 −15 26 excluded region 4 1845 17.3 1.6 2.3 15074 15380 −293733 + AluJr SINE/Alu 1 305 −7 27 6 1568 15.0 10.5 1.0 15388 15653 −293460 + AluJb SINE/Alu 1 291 −21 28 352 26.1 6.5 2.0 15654 15791 −293322 + MIR3 SINE/MIR 35 178 −30 29 689 11.4 0.0 0.0 16242 16346 −292767 C L1MB5 LINE/L1 0 6174 6070 30 2643 5.6 0.0 0.0 16374 16678 −292435 C AluY SINE/Alu −6 305 1 31 2125 10.7 3.8 0.3 16912 17200 −291913 C AluSq2 SINE/Alu −13 299 1 32 381 2.2 0.0 0.0 17660 17705 −291408 + (CA)n Simple 2 47 0 33 280 25.0 14.8 3.4 17883 17993 −291120 + MIR3 SINE/MIR 44 166 −102 34 2337 11.2 0.0 0.3 18230 18541 −290572 + AluSq2 SINE/Alu 1 311 −1 35 201 35.9 0.0 11.3 18752 18908 −290205 C L2c LINE/L2 −1 3386 3246 36 254 32.5 5.9 2.6 19294 19505 −289608 + L2b LINE/L2 3073 3286 −89 37 217 21.9 0.0 0.0 19530 19570 −289543 + (CA)n Simple 2 42 0 38 2506 8.1 0.0 0.0 19616 19923 −289190 C AluY SINE/Alu −3 308 1 39 639 21.8 3.1 2.2 19966 20118 −288995 + MIRb SINE/MIR 6 162 −106 40 probe 3 639 21.8 3.1 2.2 19966 20118 −288995 + MIRb SINE/MIR 6 162 −106 40 0 1555 15.4 8.4 2.6 20654 20974 −288139 C MER44A DNA/TcMT 0 339 1 41 381 16.3 15.1 7.4 21186 21311 −287802 C MER5A DNA/hAT-Charlie −54 135 1 42 229 22.5 6.5 4.2 21507 21599 −287514 C X8_LINE LINE/CR1 −29 267 173 43 200 38.8 3.6 2.9 22836 22973 −286140 + MIR SINE/MIR 49 187 −75 44 1354 22.8 13.0 2.1 23166 23655 −285458 + MLT1E2 LTR/ERVL-MaLR 2 541 −86 45 399 20.9 0.0 6.0 23697 23808 −285305 C MIR SINE/MIR −75 193 97 46 excluded region 5 2288 12.0 0.7 0.0 24330 24637 −284476 C AluSx1 SINE/Alu 0 312 3 47 11 2339 9.7 0.3 0.3 25459 25758 −283355 C AluSx SINE/Alu −12 300 1 48 1409 9.1 0.0 0.0 25759 25933 −283180 C AluSq2 SINE/Alu −4 308 134 49 1785 12.8 0.0 1.6 25934 26184 −282929 C AluSx SINE/Alu −12 300 54 50 916 10.5 0.0 2.5 26186 26309 −282804 + AluSx SINE/Alu 178 298 −14 51 1897 16.1 0.7 1.0 26638 26936 −282177 C AluJr SINE/Alu −14 298 1 52 189 21.1 13.8 7.6 27056 27142 −281971 C L2a LINE/L2 −3 3423 3332 53 713 22.6 2.4 3.6 27280 27307 −281806 C AluJb SINE/Alu −144 168 141 54 1795 13.9 7.9 0.7 27308 27587 −281526 C AluJb SINE/Alu −12 300 1 55 713 22.6 2.4 3.6 27588 27728 −281385 C AluJb SINE/Alu −172 140 1 54 2417 7.8 0.0 1.7 27734 28039 −281074 C AluSc SINE/Alu −7 302 2 56 2080 14.0 1.0 1.9 28040 28353 −280760 C AluSz SINE/Alu −1 311 1 57 probe 4 200 17.6 0.0 0.0 29069 29102 −280011 + C-rich Low_Cplxty 146 179 0 58 1 2386 8.5 1.3 1.6 29863 30169 −278944 + AluSc8 SINE/Alu 1 306 −6 59 excluded region 6 2494 7.4 0.0 0.0 31175 31470 −277643 C AluSg SINE/Alu −14 296 1 60 16 886 20.8 3.0 0.5 31677 31814 −277299 + MER3 DNA/hAT-Charlie 1 142 -67 61 1112 16.3 0.0 1.8 31815 31980 −277133 C AluJo SINE/Alu −13 299 137 62 886 20.8 3.0 0.5 31981 32044 −277069 + MER3 DNA/hAT-Charlie 143 207 −2 61 396 0.0 0.0 0.0 32317 32360 −276753 + (CA)n Simple 2 45 0 63 2102 9.2 0.0 0.0 32415 32675 −276438 C AluSx3 SINE/Alu −15 297 37 64 2319 9.0 0.0 1.7 32917 33217 −275896 + AluY SINE/Alu 1 296 −15 65 2269 10.2 2.4 0.0 33230 33524 −275589 + AluSp SINE/Alu 1 302 −11 66 1969 16.6 0.0 0.3 33980 34275 −274838 C AluJb SINE/Alu −16 296 2 67 2311 8.8 0.3 2.3 34281 34585 −274528 C AluSq2 SINE/Alu −13 299 1 68 199 36.4 1.5 0.0 34736 34801 −274312 + MIRc SINE/MIR 60 126 −142 69 809 26.0 0.7 9.3 34870 34901 −274212 + MIR SINE/MIR 5 33 −229 70 1727 18.2 0.0 5.9 34902 35038 −274075 + AluSx SINE/Alu 1 136 −176 71 1897 14.9 0.0 0.4 35039 35313 −273800 + AluSx SINE/Alu 1 274 −38 72 1727 18.2 0.0 5.9 35314 35496 −273617 + AluSx SINE/Alu 137 303 −9 71 809 26.0 0.7 9.3 35497 35710 −273403 + MIR SINE/MIR 34 230 −32 70 1810 17.4 1.3 1.6 35711 36014 −273099 + AluJb SINE/Alu −9 303 1 73 809 26.0 0.7 9.3 36015 36046 −273067 + MIR SINE/MIR 231 262 0 70 670 20.9 3.3 12.7 36048 36228 −272885 + FRAM SINE/Alu 1 166 0 74 437 34.5 4.7 6.3 36250 36506 −272607 + MIRb SINE/MIR 2 254 −14 75 2289 9.9 0.0 3.9 36764 37086 −272027 + AluSx1 SINE/Alu 1 311 −1 76 2440 4.5 0.0 1.1 37090 37406 −271707 + AluY SINE/Alu 1 311 0 77 1364 10.9 0.0 0.0 37407 37581 −271532 + AluSc8 SINE/Alu 133 307 −5 78 1601 18.5 0.3 4.8 37615 37916 −271197 + AluJr SINE/Alu 2 290 −22 79 probe 5 325 27.1 8.8 10.6 38602 38717 −270396 + L2c LINE/L2 2331 2446 −973 80 2107 10.4 0.3 3.2 38718 39005 −270108 + AluSx1 SINE/Alu 1 280 −32 81 414 0.0 0.0 0.0 39006 39015 −270062 + (CAA)n Simple 3 48 0 82 325 27.1 8.8 10.6 39052 39115 −269998 + L2c LINE/L2 2447 2509 −910 80 218 28.1 9.7 3.2 39093 39298 −269815 + L2c LINE/L2 2464 2682 −737 80 excluded region 7 218 28.1 9.7 3.2 39093 39298 −269815 + L2c LINE/L2 2464 2682 −737 80 9 198 0.0 0.0 0.0 39435 39456 −269657 + (TTA)n Simple 2 23 0 83 1165 10.7 0.0 0.0 39457 39605 −269508 C AluSx SINE/Alu −27 285 137 84 1808 10.0 11.9 1.0 39609 39877 −269236 C AluSp SINE/Alu −15 298 1 85 984 11.4 0.0 0.8 39890 40020 −269093 C AluSx SINE/Alu −179 133 4 84 1982 13.2 0.3 5.6 40025 40342 −268771 C AluSz SINE/Alu −10 302 1 86 2106 14.2 0.6 0.6 40380 40690 −268423 + AluSz SINE/Alu 1 311 −1 87 460 35.3 7.3 3.8 40691 41046 −268067 + L2c LINE/L2 3015 3382 −5 80 2297 10.7 0.0 0.7 41122 41420 −267693 C AluSz SINE/Alu −15 297 1 88 205 30.4 0.0 0.0 41578 41633 −267480 + (TA)n Simple 1 56 0 89 1733 20.1 0.3 0.3 41635 41928 −267185 C AluJr4 SINE/Alu −16 296 3 90 2129 12.4 0.7 0.0 42139 42429 −266684 C AluSx SINE/Alu −16 296 4 91 2203 10.4 1.0 0.0 42431 42719 −266394 C AluSp SINE/Alu −15 298 7 92 probe 6 189 0.0 0.0 0.0 44176 44196 −264917 + (CAG)n Simple 2 22 0 93 2 2434 8.6 0.0 0.0 44364 44664 −264449 C AluY SINE/Alu −9 302 2 94 2200 10.7 1.6 1.6 44923 45230 −263883 + AluSp SINE/Alu 1 308 −5 95 804 27.1 11.1 9.7 45271 45749 −263364 C L3 LINE/CR1 −188 3911 3427 96 excluded region 8 2148 13.0 0.3 0.0 45943 46243 −262870 C AluSg SINE/Alu −7 303 2 97 6 2489 7.2 0.3 0.3 46349 46653 −262460 C AluSq2 SINE/Alu −7 305 1 98 2380 8.9 0.0 1.6 46776 47089 −262024 C AluSc SINE/Alu 0 309 1 99 413 12.9 2.7 4.2 47300 47372 −261741 + L1PA8 LINE/L1 6086 6157 −15 100 436 5.8 0.0 0.0 47373 47424 −261689 C AluSz6 SINE/Alu −12 300 249 101 198 0.0 0.0 0.0 47427 47448 −261665 + (A)n Simple 1 22 0 102 2545 6.1 0.0 0.0 47532 47826 −261287 + AluY SINE/Alu 1 295 −16 103 827 16.6 0.0 6.1 47965 48103 −261010 + FLAM_C SINE/Alu 1 131 −12 104 probe 7 2366 9.4 0.3 0.0 49470 49768 −259345 C AluSp SINE/Alu −13 300 1 105 1 21 42.9 0.0 0.0 50235 50255 −258858 + AT_rich Low_Cplxty 1 21 0 106 excluded region 9 352 36.9 5.3 1.6 50840 51026 −258087 + L1MS LINE/L1 5465 5658 −584 107 16 307 30.7 16.0 0.6 51006 51149 −527964 + L1MC LINE/L1 5649 5841 −2068 108 2314 7.3 0.0 1.8 51258 51580 −257533 + AluY SINE/Alu 1 311 0 109 2432 6.5 0.0 0.3 51642 51931 −257182 + AluSp SINE/Alu 1 289 −24 110 1598 17.3 0.3 5.7 51946 52103 −257010 C AluJb SINE/Alu −19 293 142 111 2332 9.0 0.3 1.4 52104 52403 −256710 C AluSp SINE/Alu −16 297 1 112 1569 17.0 0.3 5.7 52404 52538 −256575 C AluJb SINE/Alu −171 141 15 111 754 14.3 0.9 0.0 52591 52702 −256411 + AluJr SINE/Alu 6 118 −194 113 198 10.3 0.0 0.0 53274 53302 −255811 + (TA)n Simple 1 29 0 114 2130 12.4 0.0 0.7 53303 53592 −255521 C AluSx SINE/Alu −24 288 1 115 1263 13.1 1.1 0.0 54309 54483 −254630 + AluSx1 SINE/Alu 135 311 −1 116 514 11.2 1.6 5.1 54497 54618 −254495 + GA-rich Low_Cplxty 63 180 0 117 210 15.2 0.0 0.0 54620 54652 −254461 + A-rich Low_Cplxty 1 33 0 118 190 27.9 0.0 0.0 55008 55050 −254063 C L2c LINE/L2 −15 3372 3330 119 1334 8.6 0.0 0.0 55101 55262 −253851 C AluSx1 SINE/Alu −14 298 137 120 1447 17.3 2.4 0.8 55382 55629 −253484 + AluJb SINE/Alu 37 288 −24 121 21 39.3 0.0 0.0 56454 56481 −252632 + AT_rich Low_Cplxty 1 28 0 122 2264 11.3 0.0 1.0 56869 57169 −251944 C AluSx1 SINE/Alu −14 298 1 123 2295 9.9 0.6 0.6 57258 57570 −251543 C AluSp SINE/Alu 0 313 1 124 660 16.5 0.0 12.2 57575 57624 −251489 C FLAM_C SINE/Alu −10 123 81 125 2194 11.5 0.3 0.3 57625 57920 −251193 C AluSx1 SINE/Alu −16 296 1 126 660 16.5 0.0 12.2 57921 58007 −251106 C FLAM_C SINE/Alu −53 80 1 125 1846 11.2 10.0 0.0 58454 58743 −250370 + AluSq2 SINE/Alu 1 312 0 127 probe 8 211 30.5 3.4 0.0 59728 59786 −249327 C L2b LINE/L2 −7 3368 3308 128 3 1431 8.3 0.0 0.6 59852 60031 −249082 C AluSp SINE/Alu −133 180 2 129 1870 13.5 1.8 2.1 60059 60340 −248773 + AluJo SINE/Alu 1 281 −31 130 398 16.9 2.2 5.8 60348 60436 −248677 + FLAM_A SINE/Alu 42 127 −15 131 excluded region 10 1908 14.1 5.0 0.0 62695 62991 −246122 C AluSz SINE/Alu 0 312 1 132 4 219 26.6 7.8 0.0 63055 63118 −245995 C L2a LINE/L2 −5 3421 3353 133 2274 8.9 0.7 2.0 63394 63567 −245546 C AluSx SINE/Alu −5 307 134 134 2444 8.1 0.0 0.0 63568 63865 −245248 C AluY SINE/Alu −13 298 1 135 2274 8.9 0.7 2.0 63866 64000 −245113 C AluSx SINE/Alu −179 133 2 134 probe 9 951 10.3 0.8 0.0 64794 64919 −244194 + AluSx4 SINE/Alu 179 305 −7 136 447 25.2 3.4 0.0 65518 65636 −243477 C L1ME2z LINE/L1 −3 6441 6319 137 390 4.2 0.0 0.0 65637 65684 −243429 + (CA)n Simple 1 48 0 138 319 27.9 1.2 0.0 65785 65870 −243243 + L2c LINE/L2 3295 3381 −6 139 468 29.4 4.9 2.4 66559 66913 −242200 + L1ME4a LINE/L1 5471 5849 −275 140 excluded region 11 468 29.4 4.9 2.4 66559 66913 −242200 + L1ME4a LINE/L1 5471 5849 −275 140 29 2423 10.3 0.3 0.0 66917 67227 −241886 + AluSp SINE/Alu 1 312 −1 141 1271 20.6 1.3 7.2 67277 67586 −241527 C AluJb SINE/Alu −18 294 2 142 1136 14.8 3.9 1.1 67686 67910 −241203 C L1MB3 LINE/L1 −142 6149 5936 143 319 20.7 0.0 1.7 67920 67978 −241135 C MER66C LTR/ERV1 −133 422 365 144 637 14.4 0.0 0.0 67980 68076 −241037 C L1MB3 LINE/L1 −239 5941 5845 143 2023 12.9 0.0 3.4 68567 68869 −240244 + AluSx1 SINE/Alu 1 293 −19 145 1001 10.2 0.0 0.0 69082 69208 −239905 C AluSq SINE/Alu −11 302 176 146 1879 16.8 1.0 0.7 69264 69566 −239547 + AluJb SINE/Alu 1 304 −8 147 233 30.9 0.6 0.0 69730 69811 −239302 + MIRb SINE/MIR 64 155 −113 148 2043 11.6 0.0 0.4 69909 70185 −238928 C AluSx1 SINE/Alu −11 301 26 149 2040 15.7 0.3 0.3 74836 75147 −233966 + AluJb SINE/Alu 1 312 0 150 2323 11.2 0.0 0.0 75632 75942 −233171 + AluSz SINE/Alu 2 312 0 151 1259 12.3 0.0 0.0 75957 76126 −232987 + AluSc5 SINE/Alu 130 299 −13 152 317 18.6 11.4 0.0 76427 76496 −232617 + MIR3 SINE/MIR 125 202 −6 153 818 16.1 2.8 6.4 76513 76691 −232422 + L1PREC2 LINE/L1 5984 6156 −4 154 213 14.6 3.9 6.0 76911 76961 −232152 C L2b LINE/L2 −8 3367 3318 155 859 14.5 1.5 0.8 77008 77138 −231975 + AluSz SINE/Alu 2 133 −179 156 792 26.0 4.7 0.4 77151 77382 −231731 + MIR SINE/MIR 20 261 −1 157 1679 14.3 6.3 2.0 77567 77852 −231261 C AluJr SINE/Alu −14 298 1 158 39 73.2 0.0 1.8 77874 77905 −231208 + AT_rich Low_Cplxty 1 32 0 159 2010 11.5 1.0 3.5 77906 78201 −230912 C AluSx SINE/Alu −23 289 1 160 39 73.2 0.0 1.8 78202 78225 −230888 + AT_rich Low_Cplxty 1 24 0 161 719 20.3 0.0 0.0 78226 78343 −230770 C AluJo SINE/Alu −194 118 1 162 2399 7.0 0.3 2.0 78356 78657 −230456 C AluSp SINE/Alu −15 298 2 163 2302 11.2 0.3 0.3 78796 79106 −230007 C AluSp SINE/Alu −2 311 1 164 813 14.2 2.5 0.0 79584 79703 −229410 + AluJr SINE/Alu 1 123 −189 165 1195 11.6 0.0 3.6 79875 80047 −229066 C AluSc8 SINE/Alu −16 296 130 166 891 8.6 2.8 2.2 80061 80238 −228875 + (TA)n Simple 2 180 0 167 2249 9.9 0.7 0.0 80275 80566 −228547 C AluSx SINE/Alu −18 294 1 168 2011 15.6 0.0 0.0 80729 81029 −228084 C AluSg SINE/Alu −8 302 2 169 2222 11.8 0.3 0.0 81042 81337 −227776 C AluSz SINE/Alu −15 297 1 170 1207 21.6 6.4 5.7 81444 81606 −227507 C AluJb SINE/Alu −4 298 134 171 2190 9.2 0.0 0.3 81607 81890 −227223 C AluY SINE/Alu −12 299 17 172 2382 8.4 0.0 0.0 81894 82190 −226923 C AluSc5 SINE/Alu −15 297 1 173 1612 18.7 2.8 0.7 82193 82481 −226632 C AluJo SINE/Alu −16 296 2 174 1207 21.6 6.4 5.7 82482 82605 −226508 C AluJb SINE/Alu −169 133 2 171 2381 9.5 0.0 0.0 82721 83024 −226089 + AluSx SINE/Alu 1 304 −8 175 629 20.6 2.8 0.0 83049 83155 −225958 C FLAM_A SINE/Alu −32 110 1 176 1596 9.9 0.0 0.0 83361 83561 −225552 + AluSx SINE/Alu 1 201 −111 177 402 9.6 0.0 0.0 83562 83613 −225500 + AluSx SINE/Alu 251 302 −10 177 207 0.0 0.0 0.0 83620 83642 −225471 + (GAA)n Simple 2 24 0 178 probe 11 23 56.7 0.0 0.0 83927 83956 −225157 + AT_rich Low_Cplxty 1 30 0 179 2 756 19.5 4.0 0.6 84063 84237 −224876 C MER104 Dna/TcMar-Tc2 0 181 1 180 1710 19.9 0.0 1.0 84774 85075 −224038 C AluJr SINE/Alu −12 300 2 181 298 26.3 15.7 0.7 85233 85366 −223747 C L2a LINE/L2 0 3426 3273 182 1918 12.8 4.3 0.3 85401 85681 −223432 + AluJb SINE/Alu 18 309 −3 183 700 18.2 0.0 6.0 86439 86596 −222517 + L1M4 LINE/L1 4729 4887 −1269 184 excluded region 12 700 18.2 0.0 6.0 86439 86596 −222517 + L1M4 LINE/L1 4729 4887 −1269 184 18 2561 5.3 0.3 0.0 86599 86898 −222215 C AluY SINE/Alu −10 301 1 185 1921 12.4 6.0 1.6 86905 87203 −221910 C AluSz6 SINE/Alu 0 312 1 186 645 18.4 0.0 5.2 87205 87347 −221766 + L1M4 LINE/L1 4873 5008 −1138 184 1844 13.9 3.5 0.3 87599 87885 −221228 + AluSz SINE/Alu 1 296 −16 187 2072 10.9 3.0 1.6 87965 88268 −220845 + AluSz6 SINE/Alu 1 308 −4 188 2020 8.0 8.4 0.0 88269 88554 −220559 + AluSp SINE/Alu 1 313 0 189 249 11.9 0.0 0.0 88567 88608 −220505 + (TCTA)n Simple 1 42 0 190 1260 19.2 0.5 1.4 88609 88832 −220281 C AluJr SINE/Alu −90 222 1 191 2443 7.5 0.0 0.0 89435 89729 −219384 C AluY SINE/Alu −16 295 1 192 231 23.6 6.4 2.6 89730 89827 −219286 + Tigger10 DNA/TcMT 101 204 −1639 193 1848 18.3 0.3 0.7 89841 90140 −218973 + AluJb SINE/Alu 1 299 −13 194 836 13.2 2.5 0.0 90229 90349 −218764 + AluSz SINE/Alu 1 124 −188 195 2379 9.7 0.0 0.0 90355 90652 −218461 + AluSx SINE/Alu 1 298 −14 196 771 27.4 5.0 8.2 90653 90773 −218340 + Tigger10 DNA/TcMT 841 948 −895 197 2275 11.6 0.0 0.0 90774 91074 −218039 + AluSx SINE/Alu 1 301 −11 198 2415 7.0 0.0 0.3 91077 91407 −217706 + AluY SINE/Alu 2 311 0 199 771 27.4 5.0 8.2 91408 91630 −217483 + Tigger10 DNA/TcMT 949 1180 −663 197 2276 9.3 1.0 0.0 91631 91920 −217193 C AluSx4 SINE/Alu −18 294 2 200 771 27.4 5.0 8.2 91921 91972 −217141 + Tigger10 DNA/TcMT 1181 1229 −614 197 1010 20.2 1.6 0.0 91975 92162 −216951 + AluJr4 SINE/Alu 109 299 −13 201 217 26.7 1.6 1.6 92163 92223 −216890 + (CATATA)n Simple 5 65 0 202 2319 9.6 0.7 0.0 92336 92638 −216475 C AluSp SINE/Alu −8 305 1 203 1942 13.2 0.4 0.4 92899 93202 −215911 C AluSc8 SINE/Alu 0 312 1 204 2094 11.2 3.1 0.3 93338 93623 −215490 + AluSx1 SINE/Alu 2 295 −17 205 887 20.1 0.0 0.0 93624 93767 −215346 C AluJo SINE/Alu −32 280 137 206 252 33.6 6.9 0.0 93795 93910 −215203 + Tigger15a DNA/TcMT 530 653 −62 207 probe 12 252 33.6 6.9 0.0 93795 93910 −215203 + Tigger15a DNA/TcMT 530 653 −62 207 2 468 11.4 8.6 0.0 93927 93996 −215117 C AluSq2 SINE/Alu −13 299 224 208 395 24.4 2.5 2.5 93999 94116 −214997 C Charlie4z DNA/hAT-Charlie −46 121 4 209 2373 8.8 0.3 0.0 94759 95052 −214061 + AluSx4 SINE/Alu 2 296 −16 210 23 43.5 0.0 0.0 95358 95380 −213733 + AT_rich Low_Cplxty 1 23 0 211 258 25.6 10.1 1.2 95449 95527 −213586 C L2c LINE/L2 −16 3371 3286 212 377 18.3 9.1 7.7 95752 95905 −213208 C L1MC5 LINE/L1 −36 7925 7770 213 excluded region 13 377 18.3 9.1 7.7 95752 95905 −213208 C L1MC5 LINE/L1 −36 7925 7770 213 15 728 16.7 11.4 0.0 95916 96047 −213066 C AluJo SINE/Alu −26 286 140 214 2235 10.5 0.3 0.3 96061 96354 −212759 C AluSq2 SINE/Alu −18 294 1 215 823 23.1 9.4 1.1 96357 96637 −212476 C L1MC5 LINE/L1 −444 7571 7255 213 2036 13.5 0.0 1.0 96696 96992 −212121 + AluSx4 SINE/Alu 1 294 −18 216 2148 11.7 0.3 1.3 96996 97302 −211811 + AluSg SINE/Alu 1 304 −6 217 738 27.7 8.5 2.2 97396 97904 −211209 C L2a LINE/L2 −12 3441 2870 218 1585 12.8 0.0 20.1 97915 98272 −210841 C AluJr4 SINE/Alu −14 298 1 219 1845 13.4 4.1 2.4 98298 98588 −210525 C AluSx4 SINE/Alu −15 297 2 220 497 11.0 33.0 0.0 98722 98821 −210292 + FLAM_C SINE/Alu 1 133 −10 221 237 31.1 10.1 0.0 98916 99034 −210079 + MIR3 SINE/MIR 5 135 −73 222 2590 5.3 0.0 0.0 100020 100320 −208793 + AluYk4 SINE/Alu 1 301 −11 223 1949 8.9 3.7 2.2 100331 100600 −208513 + AluSg SINE/Alu 2 275 −35 224 2347 7.8 0.0 0.0 100630 100937 −208176 + AluY SINE/Alu 1 311 0 225 2326 10.1 0.7 0.0 100941 101248 −207865 + AluSp SINE/Alu 3 312 −1 226 590 26.8 13.0 0.5 101876 102152 −206961 C L2a LINE/L2 −2 3424 3117 227 1614 16.1 1.7 2.8 102162 102300 −206813 + AluJb SINE/Alu 1 134 −168 228 2330 9.8 0.0 3.6 102301 102617 −206496 + AluY SINE/Alu 1 306 −5 229 1614 16.1 1.7 2.8 102618 102771 −206342 + AluJb SINE/Alu 135 291 −11 228 2237 9.1 2.0 0.0 102886 103183 −205930 C AluSc5 SINE/Alu −8 304 1 230 probe 13a 270 0.0 0.0 0.0 104284 104313 −204800 + (TTTTG)n Simple 1 30 0 231 1 1650 4.5 5.5 0.0 104318 104516 −204597 C AluSx SINE/Alu −37 275 66 232 excluded region 14 8064 14.0 7.8 5.5 106203 107278 −201835 + LTR12C LTR/ERV1 3 1140 −439 233 10 2324 10.1 0.0 0.3 107279 107586 −201527 + AluY SINE/Alu 2 308 −3 234 8064 14.0 7.8 5.5 107587 108052 −201061 + LTR12C LTR/ERV1 1141 1579 0 233 939 10.0 0.0 6.1 108354 108493 −200620 C FLAM_C SINE/Alu −11 132 1 235 2397 8.1 0.0 1.6 109001 109308 −199805 C AluY SINE/Alu −7 304 2 236 790 13.7 1.6 1.6 109726 109849 −199264 C FLAM_C SINE/Alu −19 124 1 237 2100 13.8 0.3 0.0 109852 110149 −198964 C AluSz SINE/Alu −13 299 1 238 696 27.4 7.1 0.9 110153 110368 −198751 C MIRc SINE/MIR −1 267 45 239 248 31.0 6.2 0.0 110411 110523 −198590 C L1M5 LINE/L1 −747 5447 5328 240 189 7.4 0.0 0.0 110917 110943 −198170 + (TAA)n Simple 2 28 0 241 1606 7.3 0.0 0.0 111079 111269 −197844 + AluY SINE/Alu 104 294 −17 242 2148 15.1 0.0 0.0 111309 111619 −197494 C AluSz6 SINE/Alu −1 311 1 243 431 16.2 14.1 0.0 111625 111723 −197390 C MIRb SINE/MIR −67 201 89 244 327 26.0 0.0 12.2 112010 112101 −197012 + MIRc SINE/MIR 37 118 −150 245 1373 9.8 0.6 0.6 112104 112286 −196827 C AluSc SINE/Alu 0 309 127 246 2444 7.5 0.0 2.9 112288 112607 −196506 C AluY SINE/Alu 0 311 1 247 251 22.8 3.5 1.7 112610 112667 −196446 + MIR SINE/MIR 104 162 −100 245 180 29.8 18.2 1.0 112901 112988 −196125 + MER5A DNA/hAT-Charlie 68 170 −19 248 2303 12.0 0.0 0.0 113162 113470 −195643 C AluSz SINE/Alu −3 309 1 249 probe 15 804 14.4 1.6 0.0 115549 115673 −193440 + FLAM_C SINE/Alu 2 128 −15 250 1 7181 6.4 0.7 0.1 115705 116977 −192136 + L1PA5 LINE/L1 4875 6154 0 251 1884 13.3 1.9 0.4 117135 117404 −191709 + AluSz SINE/Alu 1 274 −38 252 2 180 0.0 0.0 0.0 117411 117430 −191683 + (CAAAA)n Simple 1 20 0 253 2240 12.3 1.0 0.0 117441 117749 −191364 + AluSq2 SINE/Alu 1 312 0 254 224 37.7 0.0 0.0 117758 117834 −191279 + L2 LINE/L2 458 534 −2885 255 probe 16 652 29.2 9.5 7.2 118175 118595 −190518 + LTR33B LTR/ERVL 53 482 −21 256 0 722 16.5 0.0 2.5 118599 118722 −190391 + MER21C LTR/ERVL 1 121 −817 257 2342 12.3 0.0 2.8 118771 118897 −190216 C L1PREC2 LINE/L1 0 6160 6034 258 excluded region 16 2262 9.2 2.7 0.0 118898 119189 −189924 C AluSg4 SINE/Alu −12 300 1 259 1 probe 17 2262 9.2 2.7 0.0 118898 119189 −189924 C AluSg4 SINE/Alu −12 300 1 259 1 2342 12.3 0.0 2.8 119190 119429 −189684 C L1PREC2 LINE/L1 −127 6033 5803 258 1975 21.0 10.4 1.1 119430 120051 −189062 + MER21C LTR/ERVL 111 790 −148 257 279 35.6 6.5 1.6 120054 120343 −188770 + L2c LINE/L2 3030 3349 −38 260 440 17.1 4.2 6.9 120617 120735 −188378 + MLT1M LTR/ERVL-MaLR 83 198 −474 261 excluded region 17 1069 13.8 0.0 1.3 120857 121016 −188097 + AluJo SINE/Alu 135 292 −20 262 12 28 62.9 0.0 0.0 121035 121069 −188044 + AT_rich Low_Cplxty 1 35 0 263 2240 6.4 1.1 0.0 121072 121338 −187775 + AluY SINE/Alu 3 272 −39 264 2197 11.4 0.0 0.7 121453 121749 −187364 C AluSx SINE/Alu −17 295 1 265 265 28.2 1.4 1.4 121841 121912 −187201 + MIRb SINE/MIR 197 268 0 266 503 30.5 4.4 5.3 121998 122246 −186867 + MIRb SINE/MIR 19 265 −3 267 1266 11.9 0.0 1.1 122278 122453 −186660 C AluSp SINE/Alu −13 300 127 268 726 22.5 0.0 0.0 122457 122629 −186484 + (TATATG)n Simple 4 176 0 269 23 34.8 0.0 0.0 122630 122652 −186461 + AT_rich Low_Cplxty 1 23 0 270 940 11.3 0.8 0.0 122653 122776 −186337 C AluSp SINE/Alu −188 125 1 268 26 60.6 0.0 0.0 123439 123471 −185642 + AT_rich Low_Cplxty 1 33 0 271 2378 7.4 0.0 1.0 123475 123773 −185340 + AluY SINE/Alu 1 296 −15 272 784 13.1 0.0 0.0 124275 124381 −184732 + AluSx SINE/Alu 1 107 −205 273 2735 4.2 0.0 0.0 124853 125161 −183952 C AluY SINE/Alu −2 309 1 274 2424 8.1 0.0 0.0 125836 126131 −182982 C AluY SINE/Alu −3 308 13 275 1876 10.7 1.6 5.1 126545 126728 −182385 C AluSx SINE/Alu −17 295 108 276 2573 5.1 0.0 0.0 126729 127023 −182090 C AluY SINE/Alu −15 296 2 277 1876 10.7 1.6 5.1 127024 127143 −181970 C AluSx SINE/Alu −205 107 1 276 probe 18 25 72.0 0.0 0.0 127246 127270 −181843 + AT_rich Low_Cplxty 1 25 0 278 1 240 21.1 16.9 4.0 127577 127665 −181448 + MIR3 SINE/MIR 94 193 −15 279 1262 8.1 1.7 1.1 127666 127838 −181275 + AluSp SINE/Alu 124 297 −16 280 2123 13.3 16.2 0.4 127864 128270 −180843 C LTR7C LTR/ERV1 0 471 1 281 576 20.3 3.1 3.9 128487 128614 −180499 C MER2B DNA/TcMT 0 336 210 282 excluded region 18 576 20.3 3.1 3.9 128487 128614 −180499 C MER2B DNA/TcMT 0 336 210 282 4 1973 10.5 4.9 5.6 128631 128935 −180178 C AluY SINE/Alu −8 303 1 283 1150 5.9 0.0 0.0 128936 129070 −180043 C AluSz SINE/Alu −177 135 1 284 187 33.4 7.1 9.9 129286 129324 −179789 + L2 LINE/L2 2142 2181 −1238 285 2251 10.0 0.0 1.0 129325 129624 −179489 C AluSg4 SINE/Alu −14 298 2 286 187 33.4 7.1 9.9 129625 129648 −179465 + L2 LINE/L2 2182 2192 −1227 285 1745 16.7 3.5 0.0 129649 129935 −179178 C AluJb SINE/Alu −15 297 1 287 187 33.4 7.1 9.9 129936 130109 −179004 + L2 LINE/L2 2193 2374 −1045 285 probe 19 187 33.4 7.1 9.9 129936 130109 −179004 + L2 LINE/L2 2193 2374 −1045 285 2 548 25.0 0.0 0.0 130353 130464 −178649 + MER81 DNA/hAT-Bkjk 2 113 −1 288 397 20.0 3.0 1.0 130604 130704 −178409 + LTR88b LTR/Gypsy? 722 824 −13 289 1038 18.1 0.0 0.6 130839 131004 −178109 + AluSz6 SINE/Alu 7 171 −141 290 207 0.0 0.0 0.0 131023 131045 −178068 + (CAAAAA)n Simple 2 24 0 291 1739 17.6 0.0 2.7 131144 131445 −177668 + AluJr SINE/Alu 1 294 −18 292 excluded region 19 1739 17.6 0.0 2.7 131144 131445 −177668 + AluJr SINE/Alu 1 294 −18 292 18 683 21.3 8.9 2.2 131485 131652 −177461 C MIRb SINE/MIR −35 233 55 293 290 24.9 15.2 3.1 131818 131962 −177151 + L2c LINE/L2 3225 3386 −1 294 2015 12.0 0.6 1.3 131975 132108 −177005 + AluSx SINE/Alu 1 135 −177 295 2358 8.6 0.0 3.0 132109 132421 −176692 + AluY SINE/Alu 1 304 −7 296 2015 12.0 0.6 1.3 132422 132598 −176515 + AluSx SINE/Alu 136 310 −2 295 369 16.2 0.0 2.9 132682 132751 −176362 C L1MC5 LINE/L1 −523 7438 7371 297 3496 8.6 2.0 1.4 132752 133382 −175876 + LTR15 LTR/ERV1 1 671 −4 298 378 23.8 13.4 0.5 133242 133736 −175731 C L1MC5 LINE/L1 −547 7495 7255 297 2042 13.2 0.3 0.7 133441 133736 −175377 + AluSx SINE/Alu 1 295 −17 299 2238 9.5 0.0 0.0 133740 134023 −175090 + AluSg SINE/Alu 1 284 −26 300 371 4.7 0.0 0.0 134037 134079 −175034 + AluSz6 SINE/Alu 244 286 −26 301 694 29.0 9.4 4.0 134183 134701 −174412 C L2a LINE/L2 0 3375 2870 302 1211 10.9 39.0 1.0 134705 134933 −174180 C AluSx3 SINE/Alu −14 298 1 303 651 22.9 0.8 0.0 134943 135064 −174049 C AluSz SINE/Alu −187 125 3 303 1658 16.3 4.3 2.1 135083 135358 −173755 C AluSz SINE/Alu −30 282 1 304 2301 11.2 0.3 0.0 135492 135794 −173319 + AluSx SINE/Alu 1 304 −8 305 375 28.3 11.6 1.6 135871 136110 −173003 + MIRc SINE/MIR 2 268 0 306 2136 11.4 1.0 0.7 136954 137251 −171862 + AluSc8 SINE/Alu 1 299 −13 307 2368 7.1 1.0 0.3 137253 137549 −171564 + AluSp SINE/Alu 3 301 −12 308 801 26.6 8.3 0.7 138199 138452 −170661 C L2a LINE/L2 −1 3425 3153 309 1432 15.2 6.6 0.3 138490 138606 −170507 + AluJb SINE/Alu 1 117 −195 310 195 6.9 0.0 0.0 138607 138635 −170478 + (CA)n Simple 2 30 0 311 1432 15.2 6.6 0.3 138636 138788 −170325 + AluJb SINE/Alu 118 287 −25 310 254 12.8 0.0 0.0 138793 138831 −170282 + L1ME3 LINE/L1 6124 6162 0 312 1283 15.2 0.6 4.5 138839 139162 −169951 C SVA_F Other −615 760 449 313 2029 2.1 0.0 0.0 139163 139395 −169718 + SVA_C Other 1152 1384 0 314 1528 7.5 0.0 1.5 139579 139781 −169332 C AluY SINE/Alu −13 298 99 315 3520 7.6 0.2 2.8 139782 140256 −168857 C LTR2 LTR/ERV1 0 463 1 316 7381 7.3 2.1 0.0 140257 141186 −167927 C Harleq-int LTR/ERV1 0 7847 6898 316 34120 6.3 0.8 0.3 141187 145402 −163711 C Harleq-int LTR/ERV1 −996 5900 1666 316 384 4.2 0.0 0.0 145423 145470 −163643 + L1PA3 LINE/L1 6103 6150 −5 317 637 8.0 4.9 1.9 145480 145581 −163532 C Harleq-int LTR/ERV1 −5222 1674 1570 316 5813 9.7 2.9 2.2 145595 146781 −162332 C Harleq-int LTR/ERV1 −5816 1080 1 316 3514 7.8 0.4 0.2 146783 147234 −161879 C LTR2 LTR/ERV1 −10 453 1 316 775 7.8 0.0 0.0 147235 147336 −161777 C AluY LTR/ERV1 −209 102 1 315 2256 9.6 0.3 0.7 147892 148194 −160919 + AluSp SINE/Alu 1 302 −11 318 probe 22 2246 7.9 3.5 0.0 148712 149001 −160112 C AluSg SINE/Alu −9 301 2 319 2 21 42.9 0.0 0.0 150814 150834 −158279 + GC_rich Low_Cplxty 1 21 0 320 740 14.6 0.0 6.6 151349 151478 −157635 C FLAM_C SINE/Alu −21 122 1 321 excluded region 20 2502 6.8 0.0 0.3 152355 152661 −156452 C AluY SINE/Alu −5 306 1 322 5 794 13.7 1.6 1.6 152695 152818 −156295 C FLAM_C SINE/Alu −19 124 1 323 2085 13.3 1.3 0.0 152821 153120 −155993 C AluSz SINE/Alu −8 304 1 324 563 32.8 6.6 1.5 153132 153370 −155743 C MIRc SINE/MIR −10 258 3 325 791 18.7 9.2 4.2 153566 153838 −155275 + L1MC5 LINE/L1 7642 7927 −34 326 2240 9.6 0.0 0.7 153853 154145 −154968 + AluSc8 SINE/Alu 3 293 −19 327 28 67.9 0.0 0.0 154149 154176 −154937 + AT_rich Low_Cplxty 1 28 0 328 2160 9.6 2.2 3.9 154350 154662 −154451 + AluY SINE/Alu 1 308 −3 329 probe 23 216 27.8 3.8 1.2 154848 154927 −154186 + L2a LINE/L2 3302 3383 −43 330 1 298 25.0 4.6 4.6 155156 155264 −153849 + L2b LINE/L2 3256 3364 −11 331 1947 15.3 0.3 0.7 156525 156824 −152289 + AluJb SINE/Alu 1 299 −13 332 252 27.7 8.2 5.8 156901 157034 −152079 C L1Mc LINE/L1 −2228 5654 5518 333 441 0.0 0.0 0.0 157109 157157 −151956 + (CA)n Simple 2 50 0 334 315 28.3 5.2 0.0 157159 157290 −151823 C L1M5 LINE/L1 −655 5468 5326 335 excluded region 21 813 14.2 0.0 3.5 157768 157887 −151226 C AluJo SINE/Alu −196 116 1 336 3 2245 13.2 0.0 0.0 157903 158212 150901 C AluSz SINE/Alu −2 310 1 337 958 19.8 6.9 0.9 158305 158506 150607 C AluJr SINE/Alu −12 300 87 338 probe 24 515 29.2 0.6 1.3 158572 158727 −150386 C MIR SINE/MIR −106 156 2 339 0 559 23.7 7.7 1.8 159274 159428 −149685 C Tigger16b DNA/TcMT −16 321 158 340 276 19.7 0.0 0.0 159632 159697 −149416 C L1MA9 LINE/L1 −19 6293 6228 341 1903 14.2 6.8 0.3 159698 160008 −149105 C Tigger3a DNA/TcMT 0 348 18 342 304 29.1 1.7 10.2 160014 160193 −148920 C L1MA9 LINE/L1 −93 6291 6054 341 26 69.2 0.0 0.0 160250 160275 −148838 + AT_rich Low_Cplxty 1 26 0 343 excluded region 22 30 60.0 0.0 0.0 160373 160402 −148711 + AT_rich Low_Cplxty 1 30 0 344 16 1901 16.8 0.3 0.3 160410 160707 −148406 C AluJb SINE/Alu −14 298 1 345 2429 6.6 2.3 0.0 160926 161228 −147885 + AluY SINE/Alu 1 310 −1 346 2151 12.8 0.3 1.0 161239 161543 −147570 + AluSq2 SINE/Alu 1 303 −9 347 812 17.1 0.0 1.6 161559 161687 −147426 C FLAM_A SINE/Alu −13 129 3 348 2239 11.0 0.3 1.3 161748 162056 −147057 C AluSz6 SINE/Alu −6 306 1 349 637 9.0 0.8 11.5 162165 162289 −146824 C L1MA9 LINE/L1 −33 6279 6167 350 2152 13.0 0.0 0.0 162590 162598 −146515 C AluSx SINE/Alu −12 300 2 351 853 17.8 0.0 0.0 162600 162728 −146385 C FLAM_C SINE/Alu −14 129 1 352 2348 9.8 0.0 0.0 162759 163053 −146060 C AluSc SINE/Alu −13 296 2 353 753 24.7 0.0 0.7 163054 163199 −145914 C AluJb SINE/Alu −32 280 136 354 1899 16.7 2.0 0.0 163202 163449 −145619 C AluSz6 SINE/Alu −12 300 2 355 21 67.9 0.0 0.0 163511 163538 −145575 + AT_rich Low_Cplxty 1 28 0 356 1411 15.6 1.9 12.5 163577 163884 −145229 C AluJo SINE/Alu −23 289 11 357 2314 10.8 0.0 0.0 163906 164201 −144912 C AluSx SINE/Alu −16 296 1 358 2470 9.1 0.3 0.0 164346 164653 −144460 + AluSc SINE/Alu 1 309 0 359 629 21.8 7.3 0.0 164831 164954 −144159 + AluJb SINE/Alu 4 136 −176 360 1493 17.2 4.8 2.0 164955 165244 −143869 + AluJo SINE/Alu 2 299 −13 361 2231 9.3 0.0 1.4 165251 165587 −143526 + AluSq2 SINE/Alu 1 312 0 362 probe 25 5877 8.3 2.5 6.2 166057 166719 −142394 C L1PA7 LINE/L1 −1 6153 5491 363 0 excluded region 23 5877 8.3 2.5 6.2 166057 166719 −142394 C L1PA7 LINE/L1 −1 6153 5491 363 3 2432 7.4 0.0 0.7 166720 167015 −142098 C AluY SINE/Alu −17 294 1 364 5877 8.3 2.5 6.2 167016 167038 −142075 C L1PA7 LINE/L1 −664 5490 5490 363 2296 11.5 0.0 0.0 167039 167343 −141770 C AluSx3 SINE/Alu −7 305 1 365 5877 8.3 2.5 6.2 167344 167416 −141697 C L1PA7 LINE/L1 −664 5490 5420 363 2527 8.4 0.0 0.0 167417 167725 −141388 C AluY SINE/Alu −2 309 1 366 5877 7.4 1.0 0.3 167726 168279 −140834 C L1PA7 LINE/L1 −735 5491 4870 363 probe 26 5877 7.4 1.0 0.3 167726 168279 −140834 C L1PA7 LINE/L1 −735 5491 4870 363 2 1566 16.2 8.3 0.3 169630 169907 −139206 C AluJb SINE/Alu −12 300 1 367 266 33.0 2.3 1.4 169960 170120 −138993 C MIRb SINE/MIR −96 172 5 368 1633 22.3 0.0 0.7 170506 170806 −138307 + AluJr SINE/Alu 1 299 −13 369 excluded region 24 2359 8.0 0.3 0.7 171255 171556 −137557 C AluY SINE/Alu −9 302 2 370 3 2345 8.4 0.0 1.0 171557 171854 −137259 C AluSg SINE/Alu −12 298 4 371 2440 6.5 0.0 2.6 171895 172204 −136909 C AluY SINE/Alu −9 302 1 372 probe 27 500 17.8 10.2 1.4 173641 173784 −135329 + L1MC4a LINE/L1 7729 7994 −1 373 0 excluded region 25 1743 15.8 0.3 6.0 174758 174905 −134208 + AluJb SINE/Alu 2 145 −167 374 8 2453 8.3 0.3 0.0 174906 175207 −133906 + AluSp SINE/Alu 1 303 −10 375 1743 15.8 0.3 6.0 175208 175375 −133738 + AluJb SINE/Alu 146 301 −11 374 2487 8.2 0.0 0.0 175378 175681 −133432 + AluSg7 SINE/Alu 1 304 −8 376 1773 15.8 0.3 6.0 276759 276906 −32207 + AluJb SINE/Alu 2 145 −167 377 2466 8.3 0.3 0.0 276907 277207 −31906 + AluSp SINE/Alu 1 302 −11 378 1773 15.8 0.3 6.0 277208 277375 −31738 + AluJb SINE/Alu 146 301 −11 377 2510 8.5 0.0 0.0 277378 277684 −31429 + AluSg7 SINE/Alu 1 307 −5 379 probe 29 0 excluded region 26 2477 7.4 0.0 0.0 278774 279071 −30042 + AluY SINE/Alu 1 298 −13 380 6 2212 9.4 0.3 5.3 279406 279724 −29389 + AluSp SINE/Alu 1 304 −9 381 2283 10.4 0.3 0.0 279909 280205 −28908 + AluSg SINE/Alu 1 298 −12 382 2288 9.1 0.0 0.7 280216 280501 −28612 + AluY SINE/Alu 1 284 −27 383 235 22.6 7.0 2.2 280538 280623 −28490 + L1ME4a LINE/L1 5948 6037 −87 384 1552 21.2 4.2 0.3 280624 280910 −28203 C AluJb SINE/Alu −14 298 1 385 2217 8.9 1.4 0.7 280919 281210 −27903 C AluY SINE/Alu −17 294 1 386 probe 30 288 7.0 0.0 0.0 281782 281824 −27289 + (GGA)n Simple 1 43 0 387 0 excluded region 27 2005 17.0 0.0 0.0 282404 282703 −26410 C AluSz6 SINE/Alu −11 301 2 388 1 probe 31 0 excluded region 28 2341 8.6 0.7 0.7 283434 283734 −25379 + AluSx1 SINE/Alu 1 301 −11 389 1 probe 32 excluded region 29 331 28.5 9.8 2.3 283817 283938 −25175 + MIRb SINE/MIR 18 148 −120 390 0 probe 33 328 29.2 3.2 14.3 285397 285474 −23639 + MIRb SINE/MIR 3 70 −198 392 0 excluded region 30 328 29.2 3.2 14.3 285397 285474 −23639 + MIRb SINE/MIR 3 70 −198 392 10 2457 7.7 0.0 0.3 285475 285773 −23340 C AluY SINE/Alu −13 298 1 393 328 29.2 3.2 14.3 285774 285818 −23295 + MIRb SINE/MIR 71 114 −154 392 408 34.7 8.7 2.2 285879 285923 −23190 C L2c LINE/L2 −38 3349 3305 394 1815 17.3 0.0 3.3 285924 286070 −23043 + AluJb SINE/Alu 1 145 −167 395 2404 7.7 0.3 0.3 286071 286369 −22744 + AluSc5 SINE/Alu 1 299 −13 396 1815 17.3 0.0 3.3 286370 286532 −22581 + AluJb SINE/Alu 146 301 −11 395 408 34.7 8.7 2.2 286533 286611 −22502 C L2c LINE/L2 −83 3304 3221 394 2426 8.9 0.0 0.0 286612 286903 −22210 + AluSg SINE/Alu 1 292 −18 397 408 31.6 7.5 2.4 286904 287093 −22020 C L2c LINE/L2 −167 3220 3009 394 1897 18.1 0.0 0.3 287133 287435 −21678 + AluSz6 SINE/Alu 1 302 −10 398 2477 8.5 0.7 0.0 287436 287740 −21373 + AluSg SINE/Alu 1 307 −3 399 236 28.4 6.8 6.1 287743 287888 −21225 C L2c LINE/L2 −495 2924 2778 394 2425 7.2 0.7 0.0 287918 288210 −20903 + AluSx4 SINE/Alu 5 299 −13 400 1966 14.8 0.0 0.7 288319 288601 −20512 + AluJb SINE/Alu 1 281 −31 401 198 19.2 9.4 1.8 288602 288648 −20465 C L2c LINE/L2 −823 2596 2545 394 370 33.9 7.3 3.9 288662 288761 −20352 C L2c LINE/L2 −927 2492 2386 394 1455 18.4 8.1 5.3 288762 288900 −20213 C MER2 DNA/TcMT −1 344 212 402 1649 18.9 1.0 1.7 288901 289197 −19916 C AluJr SINE/Alu −17 295 1 403 1455 18.4 8.1 5.3 289198 289390 −19723 C MER2 DNA/TcMT −134 211 3 402 probe 34 1455 18.4 8.1 5.3 289198 289390 −19723 C MER2 DNA/TcMT −134 211 3 402 0 370 31.2 4.9 4.4 289391 289699 −19414 C L2c LINE/L2 −1034 2385 2033 394 274 29.6 20.4 8.6 289992 290173 −18940 C MIRb SINE/MIR −48 220 16 404 254 16.1 1.4 10.9 290149 290218 −18895 + MIR SINE/MIR 96 159 −103 405 excluded region 31 254 16.1 1.4 10.9 290149 290218 −18895 + MIR SINE/MIR 96 159 −103 405* 1998 16.9 0.0 0.3 290222 290534 −18579 + AluJb SINE/Alu 1 312 0 406 2584 6.3 0.0 0.0 290614 290913 −18200 C AluY SINE/Alu −11 300 1 407 probe 35 25 76.1 0.0 0.0 291372 291417 −17696 + AT_rich Low_Cplxty 1 46 0 408 21 38.1 0.0 0.0 291399 291419 −17694 + AT_rich Low_Cplxty 1 21 0 409 228 6.7 0.0 0.0 293811 293840 −15273 + (CAGCC)n Simple 3 32 0 410 excluded region 32 1075 11.7 0.0 1.4 295607 295751 −13362 + FLAM_C SINE/Alu 1 143 0 411 3 2297 12.3 0.0 0.3 296215 296522 −12591 + AluSx1 SINE/Alu 1 307 −5 412 2261 8.2 0.7 0.0 296524 296803 −12310 + AluSg SINE/Alu 22 303 −7 413 probe 36 611 31.6 6.1 1.2 296940 297170 −11943 C MIRb SINE/MIR −1 267 26 414 1 796 17.6 2.3 0.0 299588 299718 −9385 C FLAM_C SINE/Alu −8 135 2 415 excluded region 33 2282 9.0 0.3 0.3 299917 300205 −8908 + AluSq4 SINE/Alu 1 289 −23 416 3 1752 16.3 2.0 1.7 300991 301290 −7823 + AluSz6 SINE/Alu 2 302 −10 417 2156 13.3 0.7 0.3 301631 301930 −7183 C AluSz6 SINE/Alu −10 302 2 418 probe 37 0 excluded region 34 1844 12.7 7.6 0.0 303366 303641 −5472 + AluSz6 SINE/Alu 1 297 −15 419 6 186 4.3 0.0 0.0 303712 303734 −5379 + (TCTG)n Simple 2 24 0 420 1799 15.9 0.0 0.7 303735 304005 −5108 C AluSx3 SINE/Alu −43 269 1 421 1627 16.8 0.6 8.1 304121 304299 −4814 C AluJb SINE/Alu −3 309 129 422 2369 10.8 0.3 0.0 304300 304604 −4509 C AluSc SINE/Alu −2 307 2 423 1627 16.8 0.6 8.1 304605 304742 −4371 C AluJb SINE/Alu −184 128 14 422 365 16.1 8.5 0.0 304786 304873 −4240 C FRAM SINE/Alu 0 133 24 424 probe 38 219 3.6 0.0 0.0 305000 305027 −4086 + (CA)n Simple 2 29 0 425 0 201 7.4 0.0 0.0 305028 305054 −4059 + (TC)n Simple 2 28 0 426 262 36.0 0.0 0.0 305840 305978 −3135 + (TGG)n Simple 1 139 0 427 excluded region 35 980 19.5 0.0 1.2 306413 306573 −2540 C AluJb SINE/Alu −18 294 134 428 9 1683 16.0 0.0 1.5 306574 306841 −2272 C AluJr SINE/Alu −14 298 35 429 1081 16.8 6.0 8.0 306893 306924 −2189 C Charlie5 DNA/hAT-Charlie −1 2623 2600 430 2498 7.1 0.0 0.0 306925 307220 −1893 + AluSg SINE/Alu 1 296 −14 431 351 0.0 0.0 0.0 307222 307290 −1853 + (TA)n Simple 2 40 0 432 1081 16.8 6.0 8.0 307261 307290 −1823 C Charlie5 DNA/hAT-Charlie −25 2599 2574 430 2429 10.1 0.0 0.0 307291 307597 −1516 C AluSg SINE/Alu −3 307 1 433 1081 16.8 6.0 8.0 307598 307634 −1479 C Charlie5 DNA/hAT-Charlie −51 2573 2537 430 1814 18.1 3.4 0.0 307635 307932 −1181 + AluJr SINE/Alu 1 308 −4 434 1081 16.8 6.0 8.0 307933 307957 −1156 C Charlie5 DNA/hAT-Charlie −88 2536 2509 430 1804 16.6 1.0 1.0 307958 308258 −855 C AluJb SINE/Alu −11 301 1 435 1081 16.8 6.0 8.0 308259 308509 −604 C Charlie5 DNA/hAT-Charlie −116 2508 2251 430 180 0.0 0.0 0.0 308538 308557 −556 + (TTG)n Simple 2 21 0 436 2319 9.2 0.0 0.3 308558 308843 −270 C AluSx SINE/Alu −25 287 3 437 26 80.0 0.0 0.0 308875 308914 −199 + AT_rich Low_Cplxty 1 40 0 438 765 15.0 4.4 0.0 308915 309027 −86 + AluJo SINE/Alu 1 118 −194 439 435 14.5 0.0 0.0 309052 309113 0 C AluSz6 SINE/Alu −13 299 238 440

TABLE 4 Total Alu sequences in 11 (10.5%) probes Total Alu sequences in exlcuded 93 (89.4%) regions position in query % % % sequence (hg18) matching score div. del. ins. begin end (left) + repeat Excluded region 1 Probe 1 398 34.5 9.7 1.3 240 456 −172044 C L3 Excluded region 2 2477 7.0 0.6 1.0 2534 2845 −169655 + AluY 2391 8.5 0.0 2.3 2948 3254 −169246 + AluSg Probe 2 21 42.9 0.0 0.0 4058 4078 −168422 + AT_rich 181 13.3 0.0 0.0 5187 5216 −167284 C L2b 21 53.6 0.0 0.0 5344 5371 −167129 + AT_rich 25 44.0 0.0 0.0 6259 6283 −166217 + AT_rich 36 69.4 0.0 0.0 6261 6296 −166204 + AT_rich 300 32.4 7.6 6.2 6346 6569 −165931 C L2c Excluded region 3 2134 12.3 3.6 0.3 7463 7763 −164737 C AluSp 4581 12.2 3.9 2.7 7764 8038 −164462 + Tigger1 2268 12.5 0.0 0.0 8039 8350 −164150 C AluSz 4581 12.2 3.9 2.7 8351 8579 −163921 + Tigger1 2110 12.2 0.4 0.4 8580 8896 −163604 + AluSc 4581 12.6 5.9 2.5 8897 9223 −163277 + Tigger1 Probe 3a 4581 12.6 5.9 2.5 8897 9223 −163277 + Tigger1 722 28.2 6.0 0.9 9919 10136 −162364 C MIRb 566 16.8 1.6 2.4 11054 11181 −161319 + L1MB8 216 15.8 0.0 0.0 11954 11991 −160509 + T-rich Excluded region 4 Probe 3b 1039 34.0 8.2 3.8 14509 15076 −157424 C L2b 580 10.9 8.9 0.0 15077 15177 −157323 + L1MB4 1039 29.2 11.7 4.9 15178 15625 −156875 C L2b 392 34.2 7.0 0.0 15699 15856 −156644 + MER5B 260 27.0 2.2 1.1 16498 16587 −155913 + MER5B 356 35.0 9.7 1.8 16639 17148 −155352 + L2b Excluded region 5 356 35.0 9.7 1.8 16639 17148 −155352 + L2b Probe 3c 582 29.9 8.9 3.0 17310 18031 −154469 + L2b 570 21.9 5.8 0.6 18054 18209 −154291 + MER5A1 615 26.7 6.3 7.5 18211 18297 −154203 + L2b 463 12.4 0.0 0.0 18298 18386 −154114 C L1PB1 615 26.7 6.3 7.5 18387 18553 −153947 + L2b 616 28.0 8.3 2.9 18583 18810 −153690 C MIR 251 27.6 7.8 4.5 18895 19023 −153477 + L2b 180 24.4 18.9 0.9 19184 19278 −153222 + L2b 288 25.5 5.2 0.0 19430 19517 −152983 + MIR 409 20.3 0.9 13.5 20554 20661 −151839 + MER20 Excluded region 6 2283 10.6 0.0 0.7 20878 21178 −151322 C AluSx1 2650 5.7 0.0 0.0 21294 21593 −150907 C AluYk4 411 30.1 0.0 0.0 21609 21711 −150789 C MIR 271 27.3 6.5 0.0 21747 21823 −150677 + L1MEg 1322 24.0 7.1 2.2 21910 22707 −149793 + L1MEg 2394 10.8 0.0 0.0 22717 23021 −149479 + AluSx 367 22.0 15.0 5.0 23105 23289 −149211 + L1MEg 2251 12.5 1.6 0.0 23290 23594 −148906 + AluSx1 367 23.5 14.9 3.8 23595 23754 −148746 + L1MEg 21 66.7 0.0 0.0 23863 23883 −148617 + AT_rich 2312 9.8 0.0 0.0 23884 24168 −148332 C AluSg4 354 27.4 23.6 0.1 24296 24462 −148038 + MIRb 2271 11.0 0.0 0.3 25061 25359 −147141 C AluSq2 204 31.0 5.5 4.3 25745 25835 −146665 + L2c 189 38.0 1.8 2.7 26973 27083 −145417 + L2 3579 15.7 3.5 1.5 28391 28663 −143837 + L1MA9 2204 10.2 0.0 1.4 28664 28973 −143527 + AluSx 3579 15.7 3.5 1.5 28974 29408 −143092 + L1MA9 2260 11.5 0.0 1.9 29420 29733 −142767 C AluSx 388 29.1 18.1 0.4 30060 30252 −142248 + MIRb 2247 9.7 0.3 0.7 30637 30936 −141564 + AluSp Probe 5 467 24.0 10.4 0.0 32206 32359 −140141 C MER3 637 15.5 13.4 4.7 32864 32983 −139517 C Charlie1a Excluded region 7 637 15.5 13.4 4.7 32864 32983 −139517 C Charlie1a 2301 10.8 0.0 0.3 32984 33289 −139211 + AluSz 637 16.9 15.4 3.0 33290 33571 −138929 C Charlie1a 594 21.1 7.8 0.0 33607 33772 −138728 C Charlie1a 1745 21.7 7.6 1.8 33787 34341 −138159 C Charlie1a 2280 10.4 1.0 0.0 34508 34805 −137695 C AluSc8 25 69.2 0.0 0.0 34861 34899 −137601 + AT_rich Probe 6 551 28.8 9.0 2.0 35403 35590 −136910 + MIRb 346 34.6 12.2 4.0 35890 36193 −136307 C L2c 243 37.6 5.5 5.5 36411 36666 −135834 + L2c 186 15.2 15.2 0.0 36661 36706 −135794 C L2a 278 36.5 4.1 0.8 36911 37059 −135441 + MER5B 232 39.2 2.9 0.0 37056 37157 −135343 C L2c 293 29.1 12.7 9.0 37286 37553 −134947 C L2c 22 59.1 0.0 0.0 37814 37835 −134665 + AT_rich 1767 14.8 2.6 0.3 38038 38350 −134150 C L1MC2 2581 4.4 10.9 0.0 38351 38783 −133717 C MER9a3 2503 12.5 5.4 0.2 38790 39214 −133286 C L1MC2 Excluded region 8 2503 12.5 5.4 0.2 38790 39214 −133286 C L1MC2 2575 6.6 0.0 0.3 39220 39520 −132980 C AluY Probe 7 447 30.7 12.8 1.3 40106 40462 −132038 C L2a 1324 19.2 10.7 1.0 40694 40974 −131526 C AluJr Excluded region 9 2608 5.3 1.3 0.0 41606 41907 −130593 C AluY 1898 14.0 0.4 0.0 43234 43497 −129003 + AluSx 2028 8.5 0.4 1.2 43498 43755 −128745 + AluY 1289 15.4 0.4 8.1 43837 44089 −128411 C AluJb 1897 13.9 0.0 0.0 44300 44565 −127935 C AluSx1 311 17.9 0.0 1.5 44716 44783 −127717 + MER53 491 14.9 0.0 1.1 44783 44870 −127630 + MER53 480 14.4 4.8 11.0 45770 45894 −126606 C MER44D 1057 7.7 1.6 2.7 45879 46064 −126436 C MER44D 2405 12.7 5.6 1.2 46064 46728 −125772 C Tigger7 919 18.1 0.0 0.0 46776 46930 −125570 C MER44D 1210 14.2 11.8 0.8 47131 47342 −125158 C AluSx 967 18.1 0.0 0.0 47500 47648 −124852 + AluJb 208 22.0 1.1 6.0 47867 47953 −124547 + (TATG)n 4691 7.6 0.2 0.6 49683 50307 −122193 C L1PA10 1758 20.7 0.7 0.0 50462 50766 −121734 + AluJr4 2343 10.9 0.0 0.3 51130 51431 −121069 + AluSz 1741 18.6 1.4 0.3 51949 52244 −120256 C AluJo Probe 11 Excluded region 2443 0.4 0.0 0.8 57693 57950 −114550 + AluYa5 10 203 29.1 9.0 3.8 57957 58056 −114444 + MIRc 2301 9.7 1.0 0.3 58059 58356 −114144 + AluSx 219 18.6 3.1 15.8 58361 58424 −114076 + MIR 1903 12.7 4.4 9.5 58558 58831 −113669 C Tigger3a 2336 9.7 0.0 1.0 58832 59130 −113370 + AluSx 1903 12.7 4.4 9.5 59131 59220 −113280 C Tigger3a Probe 12 1903 12.7 4.4 9.5 59131 59220 −113280 C Tigger3a 270 39.8 0.0 0.0 60002 60119 −112381 + L4 180 11.1 0.0 0.0 60235 60261 −112239 + (A)n 474 10.8 9.2 0.0 60778 60842 −111658 C AluSq10 612 13.2 0.9 0.0 60849 60962 −111538 C Charlie1a 1915 18.2 4.9 0.7 60965 61374 −111126 C Charlie1a 321 29.3 5.9 2.1 61403 61538 −110962 C Charlie1a 1905 12.3 7.7 1.4 61652 61988 −110512 C Tigger4b 656 22.7 6.7 8.5 62213 62511 −109989 C L1MC4a 309 32.5 6.3 3.3 63088 63262 −109238 C MIRc 307 26.2 21.7 1.0 63277 63442 −109058 + HAL1 820 26.3 16.0 3.2 63465 64265 −108235 + HAL1 744 23.8 8.6 6.5 64278 64682 −107818 + HAL1 646 29.9 9.2 1.7 64710 64981 −107519 + HAL1 Excluded region 646 29.9 9.2 1.7 64710 64981 −107519 + HAL1 11 2221 11.7 2.0 0.0 65009 65307 −107193 + AluSz6 741 28.5 17.7 5.0 65308 65642 −106858 + HAL1 1932 12.4 0.4 0.0 65643 65900 −106600 + AluSx 741 25.5 7.2 8.2 65901 66135 −106365 + HAL1 513 26.8 6.3 2.2 66162 66382 −106118 + HAL1 226 27.4 8.6 9.6 66385 66535 −105965 + HAL1 2516 7.3 0.0 1.3 66536 66850 −105650 + AluY 226 27.4 8.6 9.6 66851 66926 −105574 + HAL1 4820 10.2 2.1 0.0 66927 67600 −104900 + LTR12_(—) 226 27.4 8.6 9.6 67601 67698 −104802 + HAL1 2139 11.2 0.0 0.0 67853 68168 −104332 C AluY Probe 13 460 25.0 6.8 1.9 69115 69261 −103239 + L2a 850 28.6 3.9 2.3 69391 69648 −102852 + L2a 345 23.9 19.3 1.4 69670 69788 −102712 + L2a 327 31.5 8.0 3.0 69875 70100 −102400 C L2 Excluded region 2153 8.9 2.0 1.0 71648 71776 −100724 + AluSx 12 225 0.0 0.0 0.0 71777 71801 −100699 + (TAAA)n 2153 8.9 2.0 1.0 71802 71965 −100535 + AluSx 2223 8.1 0.0 9.2 72116 72437 −100063 C AluSp Probe 14 967 25.5 2.0 3.7 73109 73356 −99144 C MIR Excluded region 2433 9.2 0.0 0.3 74262 74565 −97935 + AluSx1 13 1011 11.4 0.0 0.7 74578 74717 −97783 + AluJb 2204 12.2 0.0 0.3 74720 75007 −97493 + AluSx 2390 11.0 0.7 0.0 75008 75315 −97185 + AluSx 1873 27.2 6.0 3.0 75901 76439 −96061 C L2a 2284 9.4 1.4 0.0 76440 76725 −95775 C AluSx 1873 25.9 6.3 2.2 76726 77867 −94633 C L2a Probe 15 1873 25.9 6.3 2.2 76726 77867 −94633 C L2a 24 54.8 0.0 0.0 77993 78023 −94477 + AT_rich 1987 14.5 0.7 2.3 78087 78396 −94104 C AluJr 654 26.9 11.1 3.8 80306 80775 −91725 C HAL1 366 24.7 22.2 0.4 80915 81145 −91355 C HAL1 Excluded region 366 24.7 22.2 0.4 80915 81145 −91355 C HAL1 14 362 14.3 0.0 0.0 81186 81241 −91259 C AluJo 810 18.7 0.0 0.0 81247 81369 −91131 C AluJo 2337 10.8 1.0 0.0 81439 81745 −90755 C AluSq2 222 12.8 0.0 0.0 81790 81828 −90672 + (T)n 645 22.8 3.0 3.0 81861 82095 −90405 C HAL1 2246 12.8 0.0 0.0 82608 82904 −89596 + AluSz 870 26.0 8.8 4.5 82945 83220 −89280 + L1MC5 2237 11.4 0.0 0.7 83221 83518 −88982 + AluSx1 870 26.0 8.8 4.5 83519 83591 −88909 + L1MC5 1689 17.8 3.1 2.0 83592 83884 −88616 + AluJb 870 23.0 4.9 4.9 83885 84043 −88457 + L1MC5 2385 8.7 0.0 0.3 84076 84374 −88126 C AluSx3 361 24.7 11.5 6.8 84442 84667 −87833 C HAL1 2526 7.4 0.3 0.0 84867 85175 −87325 C AluSg4 524 30.4 1.8 0.6 85327 85495 −87005 C HAL1 510 25.4 7.2 6.6 85541 85640 −86860 + MIR 2302 10.3 0.0 0.0 85641 85941 −86559 C AluSx1 510 25.4 7.2 6.6 85942 86021 −86479 + MIR 1959 12.4 5.7 0.0 86679 86960 −85540 C AluSq2 3783 12.4 2.8 0.3 87785 88389 −84111 C Tigger1 2326 9.8 6.7 0.8 88390 88749 −83751 C THE1D 6464 20.4 3.7 4.3 88750 89064 −83436 C THE1D-int 1687 11.7 0.4 0.4 89065 89294 −83206 C AluSz6 2204 13.9 0.0 0.0 89295 89603 −82897 + AluSg 6464 20.4 3.7 4.3 89604 90942 −81558 C THE1D-int 2155 11.9 7.3 1.1 90947 91303 −81197 C THE1D 2716 11.2 3.1 1.9 91308 91627 −80873 C Tigger1 2474 7.4 0.3 0.0 91628 91926 −80574 C AluSp 2716 11.2 3.1 1.9 91927 92061 −80439 C Tigger1 691 18.9 2.0 4.8 92060 92209 −80291 C Tigger1 2112 13.6 0.7 0.3 92309 92610 −79890 + AluSz 23 65.2 0.0 0.0 93071 93093 −79407 + AT_rich 259 25.2 8.8 1.4 93163 93299 −79201 + Charlie16a 2340 9.7 0.7 0.0 93378 93675 −78825 + AluSq2 Probe 18 202 33.9 10.4 2.4 94305 94419 −78081 + MIR3 206 12.9 0.0 0.0 94740 94770 −77730 + (TTTA)n 615 27.6 3.3 3.8 94907 95117 −77383 + MIR Excluded region 323 25.3 7.1 7.8 96452 96602 −75898 C HAL1b 15 2395 10.5 0.0 0.0 96603 96907 −75593 C AluY 323 25.3 7.1 7.8 96908 97051 −75449 C HAL1b Probe 19 323 25.3 7.1 7.8 96908 97051 −75449 C HAL1b 1346 25.5 13.0 3.7 97232 97965 −74535 C L2a 795 20.8 10.2 0.0 97979 98175 −74325 C L2a 1175 5.3 0.0 0.0 98188 98319 −74181 C AluY 957 25.0 3.7 5.0 98323 98646 −73854 C L2a 1822 28.0 5.5 2.8 98660 99147 −73353 C L2a Excluded region 1822 28.0 5.5 2.8 98660 99147 −73353 C L2a 16 2307 7.8 3.8 0.0 99148 99440 −73060 + AluY 1822 28.8 8.3 1.8 99441 100520 −71980 C L2a Probe 20 1822 28.8 8.3 1.8 99441 100520 −71980 C L2a 229 9.1 0.0 0.0 100540 100583 −71917 C L1MA1 Excluded region 1871 12.6 0.0 0.0 102237 102490 −70010 + AluSx 17 Probe 21 236 24.6 4.5 2.9 102761 102827 −69673 C HAL1b 1602 16.4 3.7 0.3 102909 103217 −69283 C MLT1C 7752 5.3 1.0 0.2 103218 104175 −68325 + LTR13A Excluded region 7752 5.3 1.0 0.2 103218 104175 −68325 + LTR13A 18 1602 16.4 3.7 0.3 104176 104189 −68311 C MLT1C 1941 15.5 0.3 0.7 104190 104485 −68015 C AluSx3 1279 12.0 10.2 1.1 104490 104734 −67766 + MER47A Probe 22a 1279 12.0 10.2 1.1 104490 104734 −67766 + MER47A 1976 26.4 3.6 4.5 104810 105732 −66768 C L1MDa 298 16.3 0.0 0.0 105741 105789 −66711 + MER47A 181 32.9 3.5 2.3 106217 106303 −66197 + L2 667 17.2 9.0 0.0 106378 106499 −66001 + AluJr 584 28.8 7.0 1.0 106933 107118 −65382 C MIRb 979 25.1 18.2 0.2 107288 107655 −64845 C LTR16 Excluded region 19 Probe 22b 850 11.8 48.0 1.0 108472 108675 −63825 + AluSz 2071 22.6 7.5 3.2 108679 109832 −62668 C L1MC4a 1300 27.4 6.7 5.3 109826 110557 −61943 C L1MC4a 503 25.1 17.0 0.4 111505 111716 −60784 C MIR 26 76.9 0.0 0.0 111823 111848 −60652 + AT_rich 25 48.0 0.0 0.0 111826 111850 −60650 + AT_rich Excluded region 2266 11.9 0.0 0.7 112029 112338 −60162 C AluSz6 20 434 30.8 9.8 1.8 112397 112439 −60061 C MIRc 347 21.8 1.3 0.0 112440 112517 −59983 + MADE2 434 30.8 9.8 1.8 112518 112678 −59822 C MIRc 709 17.2 7.0 5.1 113509 113565 −58935 C MIR 1081 17.9 1.0 2.0 113566 113770 −58730 C MER6B 709 17.2 7.0 5.1 113771 113884 −58616 C MIR 922 13.4 0.0 0.8 115087 115220 −57280 + FLAM_C 2194 12.4 0.0 0.3 115855 116153 −56347 C AluSx 21 52.4 0.0 0.0 116662 116682 −55818 + AT_rich 228 22.7 0.0 0.0 118269 118312 −54188 C MARNA 334 29.6 11.7 2.5 118335 118514 −53986 C MARNA 258 28.7 4.7 4.7 119667 119816 −52684 C MER5A1 2160 12.5 0.0 0.0 121296 121598 −50902 + AluSz6 2590 4.8 0.3 2.6 121961 122276 −50224 C AluY 2312 9.6 0.3 1.0 122525 122837 −49663 C AluSq2 Probe 25 383 25.5 1.0 1.0 124840 124938 −47562 + L3 314 31.5 4.2 0.7 124992 125135 −47365 + MIRc 347 26.4 16.3 1.0 125363 125534 −46966 + L3 274 30.5 0.9 3.8 125573 125681 −46819 C L2c 501 32.6 2.8 3.6 125939 126189 −46311 + L3 399 25.0 5.7 0.2 126418 126549 −45951 C MLT1H1 24 45.8 0.0 0.0 127392 127415 −45085 + AT_rich 283 26.2 12.5 0.9 127944 128047 −44453 C L1MC5 327 26.4 0.0 0.0 128140 128230 −44270 C L1MC5 Excluded region 327 26.4 0.0 0.0 128140 128230 −44270 C L1MC5 21 504 29.0 6.4 3.1 128273 128412 −44088 C L1MC4 2235 10.0 0.3 4.5 128413 128733 −43767 + AluSz6 504 29.0 6.4 3.1 128734 128841 −43659 C L1MC4 27 40.7 0.0 0.0 128958 128984 −43516 + AT_rich 2216 10.3 0.0 0.7 129002 129293 −43207 C AluSx1 26 69.2 0.0 0.0 129304 129329 −43171 + AT_rich 716 29.2 6.6 2.7 129439 129758 −42742 C L1MC4 284 25.5 7.7 12.0 129803 129944 −42556 C L1ME4a 2477 8.5 0.0 0.0 129945 130249 −42251 C AluSx 284 25.5 7.7 12.0 130250 130445 −42055 C L1ME4a Probe 26 348 38.5 0.5 2.2 130725 130910 −41590 C MIRb 494 23.5 3.3 1.6 130919 131039 −41461 C L1M6 379 28.8 9.6 4.4 131119 131336 −41164 C MLT1J 22 63.6 0.0 0.0 131455 131476 −41024 + AT_rich 559 27.4 4.7 5.1 131889 132146 −40354 + L2a 350 23.1 2.6 0.0 132152 132229 −40271 C L1ME5 443 28.0 21.4 3.8 132249 132461 −40039 C MIR 269 25.0 12.0 0.7 132474 132606 −39894 C L1M5 582 25.6 0.8 0.0 132696 132828 −39672 + L2a Excluded region 2247 9.0 0.0 0.0 132904 133181 −39319 C AluSg 22 Probe 27 2247 9.0 0.0 0.0 132904 133181 −39319 C AluSg 2851 6.5 2.2 0.3 133284 133639 −38861 + THE1C 10891 9.9 3.9 0.6 133640 135167 −37333 + THE1C-int 2549 7.5 2.2 4.5 135168 135307 −37193 + THE1C Excluded region 2549 7.5 2.2 4.5 135168 135307 −37193 + THE1C 23 2027 12.1 0.0 8.5 135308 135638 −36862 C AluSx1 2549 7.5 2.2 4.5 135639 135862 −36638 + THE1C 256 26.8 7.8 2.7 136283 136424 −36076 C L1M6B 2419 8.7 0.0 0.7 136753 137063 −35437 C AluSq2 Probe 28a 289 30.0 4.7 5.4 137189 137336 −35164 C L2a 258 29.4 6.7 1.8 137612 137715 −34785 + MIRb 397 25.0 3.8 2.5 139471 139630 −32870 C Charlie18a 1647 17.7 2.4 4.0 139631 140006 −32494 + L1MB4 458 5.7 0.0 0.0 140640 140692 −31808 C AluYb8 245 20.4 2.0 0.0 140696 140744 −31756 C L1M5 360 20.5 13.3 0.0 141105 141238 −31262 C L1ME4a Excluded region 604 23.5 13.9 0.4 141588 141796 −30704 C MIRc 24 355 33.1 1.8 3.6 141846 142014 −30486 C MIR3 290 30.1 1.1 0.0 142104 142196 −30304 C MIR3 245 23.2 11.5 6.1 142805 142882 −29618 C L2c 189 7.4 0.0 0.0 143821 143847 −28653 + (CTGGGG)n 24 54.2 0.0 0.0 144054 144077 −28423 + GC_rich 183 8.0 0.0 0.0 144078 144102 −28398 + (CTG)n 1181 17.2 11.5 1.5 145589 145671 −26829 + MER33 2001 15.5 0.0 0.3 145672 145974 −26526 + AluJr 1181 17.2 11.5 1.5 145975 146185 −26315 + MER33 188 32.9 7.8 1.1 146389 146554 −25946 C L2 247 23.3 8.6 4.0 146683 146808 −25692 + L2c 2357 7.8 0.3 0.0 146879 147193 −25307 + AluSp 295 29.2 6.9 0.0 147406 147535 −24965 + HAL1 793 22.6 5.8 4.9 147869 148110 −24390 C MER46C 1758 10.8 0.0 0.4 148122 148352 −24148 C AluJb 722 16.0 7.9 7.5 148393 148639 −23861 + L1MB2 298 22.6 0.0 0.0 148651 148712 −23788 C MER46C 2096 9.5 4.7 1.6 149417 149712 −22788 + AluSx1 2301 9.8 0.9 2.2 149713 150028 −22472 + AluSq 264 29.2 8.3 12.8 150088 150137 −22363 C MIRb 2099 11.0 0.3 7.2 150138 150465 −22035 C AluSx 266 27.9 6.0 7.6 150466 150634 −21866 C MIRc 278 21.4 15.0 4.8 151220 151310 −21190 + L2a 2280 10.7 0.0 0.0 151311 151601 −20899 C AluSx1 278 21.4 15.0 4.8 151602 151622 −20878 + L2a 28 68.6 0.0 0.0 152478 152512 −19988 + AT_rich 2204 11.1 1.3 0.0 152585 152906 −19594 + AluSx 2129 11.3 0.0 0.7 152925 153250 −19250 C AluSz Probe 29 1328 11.5 3.0 4.3 154064 154300 −18200 C L1MA6 1331 9.1 0.5 0.0 154301 154486 −18014 + L1MA6 1253 11.9 0.0 0.0 154521 154688 −17812 + AluSp 186 4.3 0.0 0.0 154690 154712 −17788 + (CA)n 505 17.1 1.7 4.4 155541 155656 −16844 C Charlie4z Excluded region 2345 9.2 0.0 4.8 155799 156123 −16377 + AluSg4 25 2161 10.1 2.1 0.0 156545 156830 −15670 C AluSx 2127 12.2 0.0 1.7 156920 157222 −15278 C AluSz 2272 9.2 0.0 1.4 157475 157817 −14683 + AluSx 2219 3.4 2.7 0.0 157830 157956 −14544 + AluY 369 0.0 0.0 0.0 157957 157997 −14503 + (TAAA)n 2219 3.4 2.7 0.0 157998 158132 −14368 + AluY Probe 30 2231 12.0 0.3 0.7 160325 160633 −11867 C AluSx1 1987 14.8 0.3 5.8 160810 161034 −11466 C Tigger3a 1922 13.6 0.0 0.7 161035 161313 −11187 + AluSx 270 0.0 0.0 0.0 161319 161348 −11152 + (TAAA)n 1987 14.8 0.3 5.8 161349 161461 −11039 C Tigger3a Probe 31 408 29.6 1.0 11.8 161656 161862 −10638 + MER20B 628 26.9 8.4 2.9 162861 163086 −9414 C MIR 542 30.2 3.3 0.9 163485 163698 −8802 C L2 428 34.8 16.6 1.9 164306 164914 −7586 + L3 181 19.1 4.8 0.0 165048 165089 −7411 + MIRb 879 27.8 2.1 1.3 165105 165341 −7159 + Tigger13a 450 29.4 10.1 0.0 165344 165571 −6929 + Tigger13a 460 22.3 7.1 4.4 165562 165716 −6784 + Tigger13a 308 24.3 0.0 0.0 165721 165786 −6714 + MIRb 195 36.4 1.0 1.0 165816 165915 −6585 + L3 585 27.5 20.2 0.7 166018 166396 −6104 + L1M5 Excluded region 585 27.5 20.2 0.7 166018 166396 −6104 + L1M5 26 2492 6.5 0.0 0.0 166397 166690 −5810 C AluY 1414 15.4 1.4 19.3 166699 166938 −5562 C AluJb 276 3.0 0.0 0.0 166939 166971 −5529 + (TC)n 1414 15.4 1.4 19.3 166972 167083 −5417 C AluJb 237 28.2 10.3 2.2 167084 167217 −5283 + L1M5 746 18.4 0.0 3.8 167220 167355 −5145 + FLAM_C 299 25.1 8.5 1.1 167398 167562 −4938 + L1M5 1486 16.0 0.0 3.7 167618 167867 −4633 C AluJo 771 30.1 6.1 5.2 167896 168116 −4384 + L1M5 2460 9.3 0.3 0.0 168117 168428 −4072 C AluSp 771 30.1 6.1 5.2 168429 168679 −3821 + L1M5 706 21.9 4.8 8.3 168751 169044 −3456 + L1M5 2031 12.3 1.4 0.7 169045 169336 −3164 + AluSx1 716 22.1 1.1 5.0 169349 169534 −2966 + L1M4 927 20.2 1.2 1.7 169546 169718 −2782 C FAM 2029 23.8 8.0 2.8 169720 170776 −1724 + L1M4 Probe 32 2029 23.8 8.0 2.8 169720 170776 −1724 + L1M4 1480 20.6 5.8 0.0 170776 171221 −1279 + L1M2 607 26.4 0.7 0.0 171233 171376 −1124 + L1M2b 3991 25.2 2.7 3.3 171348 172500 0 + L1M2 Excluded region 3991 25.2 2.7 3.3 171348 172500 0 + L1M2 27 position in repeat Alu repeat (left) end begin linkage seq score class/family begin end (left) id (count) Excluded region 1 0 Probe 1 398 LINE/CR1 −715 3384 3150 1 0 Excluded region 2 2477 SINE/Alu 1 311 0 2 2 2391 SINE/Alu 3 302 −8 3 Probe 2 21 Low_complexity 1 21 0 4 0 181 LINE/L2 −2 3373 3344 5 21 Low_complexity 1 28 0 6 25 Low_complexity 1 25 0 7 36 Low_complexity 1 36 0 8 300 LINE/L2 −139 3248 3022 9 Excluded region 3 2134 SINE/Alu −2 311 1 10 3 4581 DNA/TcMar- 1552 1829 −589 11 Tigger 2268 SINE/Alu 0 312 1 12 4581 DNA/TcMar- 1830 2052 −366 11 Tigger 2110 SINE/Alu 1 309 0 13 4581 DNA/TcMar- 2053 2418 0 11 Tigger Probe 3a 4581 DNA/TcMar- 2053 2418 0 11 0 Tigger 722 SINE/MIR −14 254 26 14 566 LINE/L1 6051 6177 −1 15 216 Low_complexity 143 180 0 16 Excluded region 4 0 Probe 3b 1039 LINE/L2 0 3375 2752 17 0 580 LINE/L1 6070 6179 −1 18 1039 LINE/L2 −668 2751 2301 17 392 DNA/hAT-Charlie 5 173 −5 19 260 DNA/hAT-Charlie 1 91 −87 20 356 LINE/L2 687 1265 −2154 21 Excluded region 5 356 LINE/L2 687 1265 −2154 21 0 Probe 3c 582 LINE/L2 1332 2163 −1256 21 0 570 DNA/hAT-Charlie 2 165 −1 22 615 LINE/L2 2215 2285 −1134 21 463 LINE/L1 0 6151 6063 23 615 LINE/L2 2286 2466 −953 21 616 SINE/MIR 0 262 23 24 251 LINE/L2 2618 2750 −669 21 180 LINE/L2 3029 3140 −235 21 288 SINE/MIR 108 206 −62 25 409 DNA/hAT-Charlie 6 101 −118 26 Excluded region 6 2283 SINE/Alu −13 299 1 27 9 2650 SINE/Alu −12 300 1 28 411 SINE/MIR −2 260 158 29 271 LINE/L1 117 198 −6002 30 1322 LINE/L1 667 1481 −4719 30 2394 SINE/Alu 1 305 −7 31 367 LINE/L1 1665 1878 −4246 30 2251 SINE/Alu 1 310 −2 32 367 LINE/L1 1858 2035 −4165 30 21 Low_complexity 1 21 0 33 2312 SINE/Alu −27 285 1 34 354 SINE/MIR 44 240 −28 35 2271 SINE/Alu −14 298 1 36 204 LINE/L2 3252 3343 −44 37 189 LINE/L2 2741 2850 −569 38 3579 LINE/L1 5556 5823 −489 39 2204 SINE/Alu 1 312 0 40 3579 LINE/L1 5824 6279 −33 39 2260 SINE/Alu −3 309 2 41 388 SINE/MIR 40 266 −2 42 2247 SINE/Alu 1 299 −14 43 Probe 5 467 DNA/hAT-Charlie −21 188 19 44 0 637 DNA/hAT-Charlie 0 1455 1322 45 Excluded region 7 637 DNA/hAT-Charlie 0 1455 1322 45 2 2301 SINE/Alu 1 305 −7 46 637 DNA/hAT-Charlie −134 1321 988 45 594 DNA/hAT-Charlie −590 865 687 45 1745 DNA/hAT-Charlie −804 651 67 45 2280 SINE/Alu −11 301 1 47 25 Low_complexity 1 39 0 48 Probe 6 551 SINE/MIR 8 208 −60 49 0 346 LINE/L2 −79 3308 2981 50 243 LINE/L2 2910 3165 −222 51 186 LINE/L2 −98 3328 3276 52 278 DNA/hAT-Charlie 7 153 −25 53 232 LINE/L2 −648 2771 2667 50 293 LINE/L2 −2 3385 3109 54 22 Low_complexity 1 22 0 55 1767 LINE/L1 −158 6186 5867 56 2581 LTR/ERVK 0 512 33 57 2503 LINE/L1 −471 5873 5427 56 Excluded region 8 2503 LINE/L1 −471 5873 5427 56 1 2575 SINE/Alu −11 300 1 58 Probe 7 447 LINE/L2 0 3426 2972 59 1 1324 SINE/Alu −2 310 3 60 Excluded region 9 2608 SINE/Alu −5 306 1 61 10 1898 SINE/Alu 1 265 −47 62 2028 SINE/Alu 41 296 −15 63 1289 SINE/Alu −14 298 64 64 1897 SINE/Alu −2 310 45 65 311 DNA/hAT 12 78 −115 66 491 DNA/hAT 107 193 0 67 480 DNA/TcMar- −2 703 586 68 Tigger 1057 DNA/TcMar- −79 626 444 68 Tigger 2405 DNA/TcMar- −1653 838 145 69 Tigger 919 DNA/TcMar- −549 156 2 68 Tigger 1210 SINE/Alu 0 312 78 70 967 SINE/Alu 152 300 −12 71 208 Simple_repeat 3 85 0 72 4691 LINE/L1 −11 6157 5536 73 1758 SINE/Alu 1 307 −5 74 2343 SINE/Alu 1 301 −11 75 1741 SINE/Alu −9 303 5 76 Probe 11 0 Excluded region 2443 SINE/Alu 41 296 −14 77 3 10 203 SINE/MIR 63 167 −101 78 2301 SINE/Alu 1 300 −12 79 219 SINE/MIR 200 256 −6 80 1903 DNA/TcMar- 0 348 61 81 Tigger 2336 SINE/Alu 1 296 −16 82 1903 DNA/TcMar- −288 60 1 81 Tigger Probe 12 1903 DNA/TcMar- −288 60 1 81 1 Tigger 270 LINE/RTE-X 1467 1584 −445 83 180 Simple_repeat 1 27 0 84 474 SINE/Alu −236 76 6 85 612 DNA/hAT-Charlie −26 1429 1315 86 1915 DNA/hAT-Charlie −617 838 412 86 321 DNA/hAT-Charlie −1314 141 1 86 1905 DNA/TcMar- −1 360 3 87 Tigger 656 LINE/L1 −1844 6038 5745 88 309 SINE/MIR −19 249 70 89 307 LINE/L1 42 241 −2266 90 820 LINE/L1 271 1172 −1335 90 744 LINE/L1 1215 1627 −880 90 646 LINE/L1 1667 1958 −549 90 Excluded region 646 LINE/L1 1667 1958 −549 90 4 11 2221 SINE/Alu 1 305 −7 91 741 LINE/L1 15 396 −2111 92 1932 SINE/Alu 42 300 −12 93 741 LINE/L1 397 625 −1882 92 513 LINE/L1 743 972 −1535 92 226 LINE/L1 1945 2094 −413 92 2516 SINE/Alu 1 311 0 94 226 LINE/L1 2095 2166 −341 92 4820 LTR/ERV1 1 688 0 95 226 LINE/L1 2167 2268 −239 92 2139 SINE/Alu 0 311 2 96 Probe 13 460 LINE/L2 1657 1810 −1609 97 0 850 LINE/L2 2735 2996 −423 97 345 LINE/L2 3286 3425 −1 97 327 LINE/L2 −923 2496 2260 98 Excluded region 2153 SINE/Alu 1 129 −183 99 3 12 225 Simple_repeat 2 26 0 100 2153 SINE/Alu 130 296 −16 99 2223 SINE/Alu −18 295 1 101 Probe 14 967 SINE/MIR −2 260 17 102 0 Excluded region 2433 SINE/Alu 1 303 −9 103 5 13 1011 SINE/Alu 1 139 −173 104 2204 SINE/Alu 2 288 −24 105 2390 SINE/Alu 1 310 −2 106 1873 LINE/L2 −8 3418 2826 107 2284 SINE/Alu −22 290 1 108 1873 LINE/L2 −594 2825 1505 107 Probe 15 1873 LINE/L2 −594 2825 1505 107 1 24 Low_complexity 1 31 0 109 1987 SINE/Alu −6 306 2 110 654 LINE/L1 −1 2506 2003 111 366 LINE/L1 −698 1809 1529 111 Excluded region 366 LINE/L1 −698 1809 1529 111 15 14 362 SINE/Alu −10 302 247 112 810 SINE/Alu −189 123 1 113 2337 SINE/Alu −2 310 1 114 222 Simple_repeat 1 39 0 115 645 LINE/L1 −1173 1334 1100 111 2246 SINE/Alu 1 297 −15 116 870 LINE/L1 6652 6915 −1046 117 2237 SINE/Alu 1 296 −16 118 870 LINE/L1 6916 7007 −954 117 1689 SINE/Alu 3 298 −14 119 870 LINE/L1 7008 7187 −774 117 2385 SINE/Alu −1 311 14 120 361 LINE/L1 −1433 1074 839 111 2526 SINE/Alu −2 310 1 121 524 LINE/L1 −2066 441 271 111 510 SINE/MIR 78 186 −76 122 2302 SINE/Alu −11 301 1 123 510 SINE/MIR 187 259 −3 122 1959 SINE/Alu −14 298 1 124 3783 DNA/TcMar- 0 2418 1799 125 Tigger 2326 LTR/ERVL-MaLR 0 381 1 126 6464 LTR/ERVL-MaLR 0 1651 1336 126 1687 SINE/Alu −16 296 67 127 2204 SINE/Alu 2 310 0 128 6464 LTR/ERVL-MaLR −316 1335 5 126 2155 LTR/ERVL-MaLR 0 381 3 126 2716 DNA/TcMar- −617 1801 1473 125 Tigger 2474 SINE/Alu −12 301 2 129 2716 DNA/TcMar- −946 1472 1341 125 Tigger 691 DNA/TcMar- −2271 147 2 130 Tigger 2112 SINE/Alu 1 303 −9 131 23 Low_complexity 1 23 0 132 259 DNA/hAT-Charlie 195 341 −1 133 2340 SINE/Alu 1 300 −12 134 Probe 18 202 SINE/MIR 82 205 −3 135 0 206 Simple_repeat 2 32 0 136 615 SINE/MIR 34 243 −19 137 Excluded region 323 LINE/L1 −1336 673 523 138 1 15 2395 SINE/Alu −6 305 1 139 323 LINE/L1 −1487 522 380 138 Probe 19 323 LINE/L1 −1487 522 380 138 1 1346 LINE/L2 −1 3425 2625 140 795 LINE/L2 −869 2550 2334 140 1175 SINE/Alu −179 132 1 141 957 LINE/L2 −1091 2328 2009 140 1822 LINE/L2 −1465 1954 1460 140 Excluded region 1822 LINE/L2 −1465 1954 1460 140 1 16 2307 SINE/Alu 1 304 −7 142 1822 LINE/L2 −1960 1459 259 140 Probe 20 1822 LINE/L2 −1960 1459 259 140 0 229 LINE/L1 0 6302 6259 143 Excluded region 1871 SINE/Alu 44 297 −15 144 1 17 Probe 21 236 LINE/L1 −1785 224 157 138 0 1602 LTR/ERVL-MaLR −19 448 130 145 7752 LTR/ERVK 1 966 0 146 Excluded region 7752 LTR/ERVK 1 966 0 146 1 18 1602 LTR/ERVL-MaLR −338 129 115 145 1941 SINE/Alu −16 296 2 147 1279 DNA/TcMar- 30 296 −70 148 Tigger Probe 22a 1279 DNA/TcMar- 30 296 −70 148 1 Tigger 1976 LINE/L1 −3919 2699 1780 149 298 DNA/TcMar- 307 355 −11 150 Tigger 181 LINE/L2 2804 2891 −528 151 667 SINE/Alu 1 133 −179 152 584 SINE/MIR −63 205 9 153 979 LTR/ERVL −4 434 1 154 Excluded region 0 19 Probe 22b 850 SINE/Alu 1 300 −12 155 1 2071 LINE/L1 −5 7877 6672 156 1300 LINE/L1 −1660 6222 5481 156 503 SINE/MIR −14 248 2 157 26 Low_complexity 1 26 0 158 25 Low_complexity 1 25 0 159 Excluded region 2266 SINE/Alu −1 311 4 160 5 20 434 SINE/MIR −18 250 211 161 347 DNA/TcMar 1 79 −1 162 Mariner 434 SINE/MIR −58 210 30 161 709 SINE/MIR −48 214 158 163 1081 DNA/TcMar- −3 207 5 164 Tigger 709 SINE/MIR −105 157 40 163 922 SINE/Alu 1 133 −10 165 2194 SINE/Alu −14 298 1 166 21 Low_complexity 1 21 0 167 228 DNA/TcMar- −263 323 280 168 Mariner 334 DNA/TcMar- −358 228 33 168 Mariner 258 DNA/hAT-Charlie −7 159 10 169 2160 SINE/Alu 1 303 −9 170 2590 SINE/Alu −2 309 1 171 2312 SINE/Alu −1 311 1 172 Probe 25 383 LINE/CR1 2392 2490 −1609 173 0 314 SINE/MIR 119 267 −1 174 347 LINE/CR1 2843 3040 −1059 173 274 LINE/L2 −15 3372 3267 175 501 LINE/CR1 3577 3825 −274 173 399 LTR/ERVL-MaLR −368 181 1 176 24 Low_complexity 1 24 0 177 283 LINE/L1 −36 7925 7810 178 327 LINE/L1 −396 7565 7475 178 Excluded region 327 LINE/L1 −396 7565 7475 178 3 21 504 LINE/L1 −20 8022 7869 179 2235 SINE/Alu 1 308 −4 180 504 LINE/L1 −174 7868 7766 179 27 Low_complexity 1 27 0 181 2216 SINE/Alu −22 290 1 182 26 Low_complexity 1 26 0 183 716 LINE/L1 −495 7547 7216 179 284 LINE/L1 −90 6034 5888 184 2477 SINE/Alu −7 305 1 185 284 LINE/L1 −237 5887 5710 184 Probe 26 348 SINE/MIR −35 233 51 186 0 494 LINE/L1 −4691 1805 1683 187 379 LTR/ERVL-MaLR −48 464 236 188 22 Low_complexity 1 22 0 189 559 LINE/L2 3170 3426 0 190 350 LINE/L1 −321 5873 5794 191 443 SINE/MIR −4 258 8 192 269 LINE/L1 −339 5784 5637 193 582 LINE/L2 3293 3426 0 194 Excluded region 2247 SINE/Alu −31 279 2 195 1 22 Probe 27 2247 SINE/Alu −31 279 2 195 1 2851 LTR/ERVL-MaLR 3 365 −10 196 10891 LTR/ERVL-MaLR 1 1578 −2 196 2549 LTR/ERVL-MaLR 19 160 −215 196 Excluded region 2549 LTR/ERVL-MaLR 19 160 −215 196 2 23 2027 SINE/Alu −6 306 2 197 2549 LTR/ERVL-MaLR 161 375 0 196 256 LINE/L1 −156 213 65 198 2419 SINE/Alu −3 309 1 199 Probe 28a 289 LINE/L2 −4 3422 3276 200 1 258 SINE/MIR 116 224 −44 201 397 DNA/hAT-Charlie −2 340 179 202 1647 LINE/L1 5777 6146 −34 203 458 SINE/Alu −260 58 6 204 245 LINE/L1 −453 5671 5622 205 360 LINE/L1 −7 6117 5952 206 Excluded region 604 SINE/MIR −10 258 22 207 9 24 355 SINE/MIR −23 185 20 208 290 SINE/MIR −1 207 114 209 245 LINE/L2 −20 3367 3286 210 189 Simple_repeat 6 32 0 211 24 Low_complexity 1 24 0 212 183 Simple_repeat 1 25 0 213 1181 DNA/hAT-Charlie 1 81 −243 214 2001 SINE/Alu 1 302 −10 215 1181 DNA/hAT-Charlie 82 324 0 214 188 LINE/L2 −1148 2271 2095 216 247 LINE/L2 3229 3358 −17 217 2357 SINE/Alu 1 313 0 218 295 LINE/L1 150 288 −2219 219 793 DNA/TcMar- 0 338 95 220 Tigger 1758 SINE/Alu −81 231 2 221 722 LINE/L1 5942 6178 −5 222 298 DNA/TcMar- −274 64 3 220 Tigger 2096 SINE/Alu 1 305 −7 223 2301 SINE/Alu 1 312 −1 224 264 SINE/MIR −17 251 202 225 2099 SINE/Alu −5 307 1 226 266 SINE/MIR −67 201 38 225 278 LINE/L2 3303 3405 −21 227 2280 SINE/Alu −21 291 1 228 278 LINE/L2 3406 3426 0 227 28 Low_complexity 1 35 0 229 2204 SINE/Alu 10 312 0 230 2129 SINE/Alu 0 312 1 231 Probe 29 1328 LINE/L1 −7 6293 6060 232 1 1331 LINE/L1 5791 5977 −323 232 1253 SINE/Alu 137 304 −9 233 186 Simple_repeat 2 24 0 234 505 DNA/hAT-Charlie 0 167 55 235 Excluded region 2345 SINE/Alu 1 310 −2 236 6 25 2161 SINE/Alu −20 292 1 237 2127 SINE/Alu −14 298 1 238 2272 SINE/Alu 6 312 0 239 2219 SINE/Alu 1 127 −184 240 369 Simple_repeat 2 42 0 241 2219 SINE/Alu 128 269 −42 240 Probe 30 2231 SINE/Alu −4 308 1 242 2 1987 DNA/TcMar- −20 328 106 243 Tigger 1922 SINE/Alu 1 277 −35 244 270 Simple_repeat 3 32 0 245 1987 DNA/TcMar- −243 105 2 243 Tigger Probe 31 408 DNA/hAT-Charlie 2 188 −595 246 0 628 SINE/MIR −23 239 2 247 542 LINE/L2 −745 2674 2456 248 428 LINE/CR1 655 1352 −2747 249 181 SINE/MIR 144 187 −81 250 879 DNA/TcMar- 12 250 −521 251 Tigger 450 DNA/TcMar- 342 592 −179 252 Tigger 460 DNA/TcMar- 607 765 −6 253 Tigger 308 SINE/MIR 197 262 0 254 195 LINE/CR1 1344 1443 −2656 249 585 LINE/L1 2518 2973 −3173 255 Excluded region 585 LINE/L1 2518 2973 −3173 255 6 26 2492 SINE/Alu −16 295 2 256 1414 SINE/Alu −2 300 115 257 276 Simple_repeat 2 34 0 258 1414 SINE/Alu −188 114 1 257 237 LINE/L1 2981 3118 −3028 255 746 SINE/Alu 2 132 −11 259 299 LINE/L1 3219 3395 −2751 255 1486 SINE/Alu −20 292 52 260 771 LINE/L1 3410 3626 −2520 255 2460 SINE/Alu 0 313 1 261 771 LINE/L1 3627 3886 −2260 255 706 LINE/L1 3929 4208 −1938 255 2031 SINE/Alu 1 294 −18 262 716 LINE/L1 2 180 −6362 263 927 SINE/Alu −13 172 1 264 2029 LINE/L1 188 1298 −5244 263 Probe 32 2029 LINE/L1 188 1298 −5244 263 0 1480 LINE/L1 1 472 −6377 265 607 LINE/L1 498 642 −6567 266 3991 LINE/L1 581 1642 −5207 265 Excluded region 3991 LINE/L1 581 1642 −5207 265 0 27

TABLE 5 Description of the 6 characterized large rearrangements as detected by MLPA and Molecular Combing Sample Gene MLPA status Molecular Combing Breakpoints (bp) Mechanism Mutation name Reference 1 BRCA1 Dup ex 13 6.1 ± 1.6 kb/ 38483825-38489905 Alu-Alu HR c.4186-1785_4358- Puget et al. (1999) Dup ex 13 1667dup6081 2 BRCA1 Del ex 2 40.8 ± 3.5 kb/ NBR1 38 562 663-38 562 427; Pseudogen-Alu c.-33024_80 + Puget N, 2002 Del ex 2 BRCA1 38 525 728-38 525 492 3832del36936 Am J Hum Genet 70: 858-865 3 BRCA1 Del ex 2 39.0 ± 2.6 kb/ NBR1 38 562 663-38 562 427; Pseudogen-Alu c.-33024_80 + Puget N, 2002 Del ex 2 BRCA1 38 525 728-38 525 492 3832del36936 Am J Hum Genet 70: 858-865 4 BRCA1 Dup ex 18-20 6.7 ± 1.2 kb/ 38460514-38470596 Alu-Alu HR c.5075- Staaf et al. (2008) Dup ex 18-20 1093_5277 + 2089dup10082 5 BRCA1 Del ex 15 4.1 ± 1.2 kb/ 38478177_38481174 Alu-Alu HR c.4484 + Puget et al. (1999b) Del ex 15 857_4676-1396del 6 BRCA1 Del ex 8-13 20 ± 2.8 kb/ 38,507,324-38,483,560 Alu-Alu HR c.442-1901_4358- Puget et al. (1999b) Del ex 8-13 1404del23763 All patients were previously characterized by high resolution aCGH, and the reported values were originally described by Rouleau et al (Rouleau 2007).

TABLE 6 Robustness of BRCA1 and BRCA2 signals measurement in 10 control blood donors BRCA1 - mean measured motifs length Blood donor g1b1 g2b1 g3b1 g4b1 g5b1 g6b1 g7b1 7232 8.6 10.0 13.3 16.9 19.9 9.9 16.5 7673 8.4 9.9 14.2 17.5 20.8 11.2 18.2 7639 7.7 8.8 11.5 15.3 18.0 8.4 15.0 7671 7.6 10.6 11.0 16.7 19.4 9.6 16.0 7672 8.4 10.0 13.0 16.8 20.2 9.9 17.3 An 8 7.1 8.2 12.2 14.9 18.7 8.2 15.9 An 11 8.6 9.4 11.8 16.4 20.5 9.6 17.4 An 12 8.6 11.0 12.5 17.0 20.8 11.2 18.0 An 13 8.7 9.9 13.6 17.1 20.2 9.8 17.6 An 14 8.4 9.8 12.2 16.5 20.2 9.5 17.5 μ (measured) 8.2 9.7 12.5 16.5 19.9 9.7 16.9 SD 0.5 0.8 0.9 0.8 0.9 0.9 1.0 calculated 8.5 9.5 12.3 16.5 19.7 9.3 17.7 delta 0.3 −0.2 −0.2 0.0 −0.2 −0.4 0.8 BRCA2 - mean measured motifs length Blood donor g1b2 g2b2 g3b2 g4b2 g5b2 7232 20.2 24.0 15.6 20.6 21.3 7673 22.6 24.4 16.4 22.3 22.4 7639 19.7 21.3 15.5 19.2 19.4 7671 20.7 22.3 15.9 21.3 21.3 7672 21.2 23.4 16.9 21.7 21.3 An 8 20.6 21.1 15.2 20.3 19.5 An 11 22.1 23.9 15.8 21.9 21.9 An 12 21.7 24.7 17.3 22.9 21.9 An 13 22.6 22.2 16.6 21.2 20.8 An 14 22.6 23.7 17.2 22.3 21.7 μ (measured) 21.4 23.1 16.2 21.4 21.2 SD 1.0 1.2 0.7 1.1 1.0 calculated 20.7 23.5 16.1 21.1 20.8 delta −0.7 0.4 −0.1 −0.3 −0.4

BRCA1 motifs g1b1 to b4b1 Case number g1b1 (8.5) g2b1 (9.5) g3b1 (12.3) g4b1 (16.5) BRCA1 m CV m CV m CV m CV Mutation n (kb) delta SD (%) SF n (kb) delta SD (%) SF n (kb) delta SD (%) SF SEM 95% CI Error n (kb) delta SD (%) SF SEM 95% CI Error 1 Tot 36 8.0 −0.5 1.2 15.0 2.1 36 10.1 0.6 2.4 23.8 1.9 38 11.6 −0.7 2.1 18.1 2.1 40 19.0 2.5 3.5 18.4 1.7 Dup wt 21 16.1 1.6 9.8 2.0 0.3 15.4 16.8 ex 13 mut 19 22.2 2.0 8.9 1.5 0.5 21.3 23.1 delta 6.1 4.5 7.7 1.6 2 Tot 21 8.2 −0.3 1.8 22.0 2.1 18 9.2 −0.3 1.8 19.6 2.1 17 12.0 −0.3 1.9 15.8 2.1 22 15.9 −0.6 2.2 13.8 2.1 Del ex wt 36 kb mut delta 3 Tot 23 8.8 0.3 3.1 35.1 1.9 25 11.4 1.9 3.3 28.7 1.7 25 11.6 −4.9 3.5 30.4 2.1 25 16.7 0.2 2.2 13.3 2.0 Del wt ex 2 mut 39 kb delta 4 Tot 28 8.0 −0.5 1.3 16.0 2.1 30 9.7 0.2 2.3 24.0 2.0 33 15.0 2.7 3.5 23.3 1.6 30 16.5 0.0 2.8 17.0 2.0 Dup wt 22 12.7 1.1 8.7 2.6 0.2 12.2 13.2 ex mut 11 19.4 1.2 6.2 1.7 0.4 18.7 20.1 18-20 delta 6.7 5.5 7.9 1.2 5 Tot 31 8.0 −0.5 1.0 11.9 2.1 32 9.8 0.3 1.5 15.3 1.9 33 11.7 −0.6 1.9 16.0 2.1 33 14.3 −2.2 2.3 16.1 2.3 Del wt 12 16.9 1.3 7.6 2.0 0.4 16.2 17.7 ex 15 mut 21 12.8 1.1 8.8 2.6 0.2 12.3 13.3 delta −4.1 −5.3 −2.8 1.2 6 Tot 21 8.8 0.3 0.9 10.2 1.9 22 10.8 1.3 1.9 17.6 1.9 22 11.6 −0.7 1.9 16.4 1.9 23 17.5 −2.2 4.0 23.0 2.2 Del wt 13 20.8 1.6 7.9 1.6 0.5 19.9 21.7 ex 8-13 mut 10 13.3 1.1 8.0 2.5 0.3 12.6 14.0 delta −7.5 −9.0 −5.9 1.6 BRCA1 motifs g5b1 to g7b1 g5b1 (19.7) g6b1 (9.3) g7b1 (17.7) Case number m CV m CV m CV BRCA1 Mutation n (kb) delta SD (%) SF SEM 95% CI Error n (kb) delta SD (%) SF n (kb) delta SD (%) SF SEM 95% CI Error 1 Tot 37 18.5 −1.2 2.8 15.1 2.1 34 9.1 −0.2 1.9 20.9 2.0 31 16.2 Dup ex 13 wt mut delta 2 Tot 22 19.2 −0.5 1.3 6.8 2.1 18 8.8 −0.5 2.5 28.4 2.1 20 12.3 Del ex wt 11 18.1 0.7 3.9 2.0 0.2 17.7 18.5 36 kb mut 9 8.1 1.6 19.8 4.4 0.5 7.0 9.2 delta −10.0 −11.5 8.5 1.5 3 Tot 19 19.6 −0.1 2.7 13.9 2.0 19 10.5 1.2 3.0 28.3 1.8 16 11.9 Del ex 2 wt 5 17.3 0.5 2.9 2.0 0.2 16.9 17.7 39 kb mut 11 8.7 1.2 13.8 4.1 0.4 8.0 9.4 delta −8.6 −9.8 −7.4 1.2 4 Tot 24 20 0.3 2.3 11 2 23 9.8 0.5 1.6 17 1.9 22 17.2 Dup ex 18-20 wt mut delta 5 Tot 28 19.4 −0.3 1.9 9.8 2.0 22 9.3 0.0 1.2 12.5 2.0 20 17.8 Del ex 15 wt mut delta 6 Tot 23 12.8 −3.7 5.5 43.0 2.6 21 10.5 1.2 1.9 18.1 1.8 20 18.0 0.3 2.4 13.3 2.1 Del ex 8-13 wt 13 18.3 1.3 7.1 1.8 0.4 17.6 19.0 mut 10 5.8 0.5 8.6 5.7 0.2 5.5 6.1 delta −12.5 13.5 −11.5 1.0 BRCA2 motifs g1b2 to g5b2 g1b2 (20.7) g2b2 (23.5) g3b2 (16.1) g4b2 (21.1) g5b2 (20.8) m CV m CV m CV m CV m CV case nr. n (kb) delta SD (%) SF n (kb) delta SD (%) SF n (kb) delta SD (%) SF n (kb) delta SD (%) SF n (kb) delta SD (%) SF 1 Tot 24 20.2 −0.5 2.6 12.9 2.0 25 22.2 −1.3 4.6 20.7 2.1 30 16.1 0.0 2.1 13.0 2.0 28 20.6 −0.5 2.7 13.1 2.0 27 20.7 −0.1 2.4 11.6 2.0 wt mut delta 2 Tot 31 20.2 −0.5 2.0 9.9 2.0 23 22.2 −1.3 1.0 4.5 2.1 31 15.2 −0.9 1.2 7.9 2.1 28 20.5 −0.6 1.4 6.8 2.1 23 20.9 0.1 2.1 10.0 2.0 wt mut delta 3 Tot 26 20.3 −0.4 2.2 10.8 2.0 28 22.5 −1.0 3.4 15.1 2.1 30 16.8 0.7 2.2 13.1 1.9 30 21.0 −0.1 2.5 11.9 2.0 28 20.3 −0.5 2.9 14.3 2.0 wt mut delta 4 Tot 21 21.3 0.6 3.2 15.0 1.9 23 22.0 −1.5 3.7 16.8 2.1 30 16.2 0.1 2.9 17.9 2.0 27 20.7 −0.4 1.9 9.2 2.0 19 20.9 0.1 2.6 12.4 2.0 wt mut delta 5 Tot 27 21.5 0.8 2.1 9.8 1.9 28 22.6 −0.9 2.0 8.8 2.1 29 16.6 0.5 2.2 13.3 1.9 28 22.4 1.3 2.1 9.4 1.9 23 21.3 0.5 1.4 6.6 2.0 wt mut delta 6 Tot 21 22.6 1.9 2.0 8.8 1.8 22 24.4 0.9 2.7 11.1 1.9 22 18.0 1.9 2.1 11.7 1.8 20 22.8 1.7 0.9 3.9 1.9 17 22.3 1.5 1.4 6.3 1.9 wt mut delta

TABLE 8 SEQ ID NO°   1 BRCA1-1A-F DNA Homo sapiens GGGACGGAAAGCTATGATGT SEQ ID NO°   2 BRCA1-1A-R DNA Homo sapiens GGGCAGAGGTGACAGGTCTA SEQ ID NO°   3 BRCA1-1B-F DNA Homo sapiens CCTCTGACCTGATCCCTTGA SEQ ID NO°   4 BRCA1-1B-R DNA Homo sapiens ATCAGCAACAGTCCCATTCC SEQ ID NO°   5 BRCA1-2-F DNA Homo sapiens GCCCAGACTAGTGTTTCTTAACC SEQ ID NO°   6 BRCA1-2-R DNA Homo sapiens GGCATGAGGCAGCAATTTAG SEQ ID NO°   7 BRCA1-3-F DNA Homo sapiens TCTTTGAATCTGGGCTCTGC SEQ ID NO°   8 BRCA1-3-R DNA Homo sapiens GCTGTTGCTTTCTTTGAGGTG SEQ ID NO°   9 BRCA1-4-F DNA Homo sapiens CACAGGTATGTGGGCAGAGA SEQ ID NO°  10 BRCA1-4-R DNA Homo sapiens CCTCTGTTGATGGGGTCATAG SEQ ID NO°  11 BRCA1-5-F DNA Homo sapiens TTTGGTAGACCAGGTGAAATGA SEQ ID NO°  12 BRCA1-5-R DNA Homo sapiens CAAATTATGTGTGGAGGCAGA SEQ ID NO°  13 BRCA1-6-F DNA Homo sapiens GAAGAACGTGCTCTTTTCACG SEQ ID NO°  14 BRCA1-6-R DNA Homo sapiens AAAGTCTGATAACAGCTCCGAGA SEQ ID NO°  15 BRCA1-7-F DNA Homo sapiens TTCGATTCCCTAAGATCGTTTC SEQ ID NO°  16 BRCA1-7-R DNA Homo sapiens CACAGTTCTGTGTAATTTAATTTCGAT SEQ ID NO°  17 BRCA1-8-F DNA Homo sapiens AGGGAAGGCTCAGATACAAAC SEQ ID NO°  18 BRCA1-8-R DNA Homo sapiens TGCCATAGATAGAGGGCTTTTT SEQ ID NO°  19 BRCA1-9-F DNA Homo sapiens GCCATCTTCTTTCTCCTGCT SEQ ID NO°  20 BRCA1-9-R DNA Homo sapiens TTGACCTATTGCTGAATGTTGG SEQ ID NO°  21 BRCA1-11-F DNA Homo sapiens TTTTACCAAGGAAGGATTTTCG SEQ ID NO°  22 BRCA1-11-R DNA Homo sapiens GCTTGATCACAGATGTATGTATGAGTT SEQ ID NO°  23 BRCA1-12-F DNA Homo sapiens CCCCAGGGCTTTAAAGGTTA SEQ ID NO°  24 BRCA1-12-R DNA Homo sapiens TAGGGGTGGATATGGGTGAA SEQ ID NO°  25 BRCA1-13A-F DNA Homo sapiens ACTTCTTCAACGCGAAGAGC SEQ ID NO°  26 BRCA1-13A-R DNA Homo sapiens GACAGGCTGTGGGGTTTCT SEQ ID NO°  27 BRCA1-15-F DNA Homo sapiens TATCTGCTGGCCACTTACCA SEQ ID NO°  28 BRCA1-15-R DNA Homo sapiens TCTCGAGCCTTGAACATCCT SEQ ID NO°  29 BRCA1-16-F DNA Homo sapiens CGCTCAGCTTTCATTCCAGT SEQ ID NO°  30 BRCA1-16-R DNA Homo sapiens AAACGTTCACATGTATCCCCTAA SEQ ID NO°  31 BRCA1-17-F DNA Homo sapiens CCTGGCCAGTACCCAGTAGT SEQ ID NO°  32 BRCA1-17-R DNA Homo sapiens CTGAGCCCAGAGTTTCTGCT SEQ ID NO°  33 BRCA1-18-F DNA Homo sapiens GGGCCCAAAAACCAGTAAGA SEQ ID NO°  34 BRCA1-18-R DNA Homo sapiens GGGATTGAGCGTTCACAGAT SEQ ID NO°  35 BRCA1-19-F DNA Homo sapiens GCCATCCAGTCCAGTCTCAT SEQ ID NO°  36 BRCA1-19-R DNA Homo sapiens TGCAGTTCTACCCTCCACTTG SEQ ID NO°  37 BRCA1-22-F DNA Homo sapiens CGGGTAAGTGGTGAGCTTTC SEQ ID NO°  38 BRCA1-22-R DNA Homo sapiens GACTGTCATTTAAAGGCACTTTTT SEQ ID NO°  39 BRCA1-23-F DNA Homo sapiens TGGCTAGTGTTTTGGCCTGT SEQ ID NO°  40 BRCA1-23-R DNA Homo sapiens TTCAGTGTTGCTTCTCCATTTC SEQ ID NO°  41 BRCA1-24-F DNA Homo sapiens TGTCAGACTAGCCACAGTACCA SEQ ID NO°  42 BRCA1-24-R DNA Homo sapiens AAGCGCTTCTTCATATTCTCC SEQ ID NO°  43 BRCA1-25-F DNA Homo sapiens ACCACACTCTTCTGTTTTGATGT SEQ ID NO°  44 BRCA1-25-R DNA Homo sapiens GGCACATGTACACCATGGAA SEQ ID NO°  45 BRCA1-26-F DNA Homo sapiens TTGTGTAGGTTGCCCGTTC SEQ ID NO°  46 BRCA1-26-R DNA Homo sapiens TTCAGAGAGCTGGGCCTAAA SEQ ID NO°  47 BRCA1-27-F DNA Homo sapiens GGAGGCAATCTGGAATTGAA SEQ ID NO°  48 BRCA1-27-R DNA Homo sapiens GGATCCATGATTGCTGCTTT SEQ ID NO°  49 BRCA1-28-F DNA Homo sapiens TCTCTGCTGTTTTTACAACTTTTTC SEQ ID NO°  50 BRCA1-28-R DNA Homo sapiens GGATCCATGATTGCTGCTTT SEQ ID NO°  51 BRCA1-29-F DNA Homo sapiens CCCTCTAGATACTTGTGTCCTTTTG SEQ ID NO°  52 BRCA1-29-R DNA Homo sapiens TCTGGCAGTCACAATTCAGG SEQ ID NO°  53 BRCA1-30-F DNA Homo sapiens TCCCATGACTGCATCATCTT SEQ ID NO°  54 BRCA1-30-R DNA Homo sapiens TTGAGATCAGGTCGATTCCTC SEQ ID NO°  55 BRCA1-31-F DNA Homo sapiens AAAACTCAACCCAAACAGTCA SEQ ID NO°  56 BRCA1-31-R DNA Homo sapiens CCAAGAATCACGAAGAGAGAGA SEQ ID NO°  57 BRCA1-32-F DNA Homo sapiens GACCTCATAGAGGTAGTGGAAAGAA SEQ ID NO°  58 BRCA1-32-R DNA Homo sapiens GCTCAAAGCCTTTAGAAGAAACA SEQ ID NO°  59 BRCA1-33-F DNA Homo sapiens GCACTGGGGAAAAGGTAGAA SEQ ID NO°  60 BRCA1-33-R DNA Homo sapiens CTCTTCAACCCAGACAGATGC SEQ ID NO°  61 BRCA1-34-F DNA Homo sapiens CAATACCCAATACAATGTAAATGC SEQ ID NO°  62 BRCA1-34-R DNA Homo sapiens CTGGGGATACTGAAACTGTGC SEQ ID NO°  63 BRCA1-35-F DNA Homo sapiens ATCAAGAAGCCTTCCCAGGT SEQ ID NO°  64 BRCA1-35-R DNA Homo sapiens TCCTTGGACGTAAGGAGCTG SEQ ID NO°  65 BRCA1-36-F DNA Homo sapiens TTCAGAACTTCCAAATACGGACT SEQ ID NO°  66 BRCA1-36-R DNA Homo sapiens GATGGAGCTGGGGTGAAAT SEQ ID NO°  67 BRCA1-37-F DNA Homo sapiens CGTGAGATTGCTCACAGGAC SEQ ID NO°  68 BRCA1-37-R DNA Homo sapiens CAAGGCATTGGAAAGGTGTC SEQ ID NO°  69 BRCA1-38-F DNA Homo sapiens AGAGGAATAGACCATCCAGAAGT SEQ ID NO°  70 BRCA1-38-R DNA Homo sapiens TCCTCCAGCACTAAAAACTGC SEQ ID NO°  71 BRCA2-1-F DNA Homo sapiens AAATGGAGGTCAGGGAACAA SEQ ID NO°  72 BRCA2-1-R DNA Homo sapiens TGGAAAGTTTGGGTATGCAG SEQ ID NO°  73 BRCA2-2-F DNA Homo sapiens TCTCAATGTGCAAGGCAATC SEQ ID NO°  74 BRCA2-2-R DNA Homo sapiens TCTTGACCATGTGGCAAATAA SEQ ID NO°  75 BRCA2-3a-F DNA Homo sapiens AATCACCCCAACCTTCAGC SEQ ID NO°  76 BRCA2-3a-R DNA Homo sapiens GCCCAGGACAAACATTTTCA SEQ ID NO°  77 BRCA2-3b-F DNA Homo sapiens CCCTCGCATGTATGATCTGA SEQ ID NO°  78 BRCA2-3b-R DNA Homo sapiens CTCCTGAAGTCCTGGAAACG SEQ ID NO°  79 BRCA2-3c-F DNA Homo sapiens TGAAATCTTTTCCCTCTCATCC SEQ ID NO°  80 BRCA2-3c-R DNA Homo sapiens AGATTGGGCACATCGAAAAG SEQ ID NO°  81 BRCA2-5-F DNA Homo sapiens GGTCTTGAACACCTGCTACCC SEQ ID NO°  82 BRCA2-5-R DNA Homo sapiens CACTCCGGGGGTCCTAGAT SEQ ID NO°  83 BRCA2-6-F DNA Homo sapiens TCTTTAACTGTTCTGGGTCACAA SEQ ID NO°  84 BRCA2-6-R DNA Homo sapiens TGGCTAGAATTCAAAACACTGA SEQ ID NO°  85 BRCA2-7-F DNA Homo sapiens TTGAAGTGGGGTTTTTAAGTTACAC SEQ ID NO°  86 BRCA2-7-R DNA Homo sapiens CCAGCCAATTCAACATCACA SEQ ID NO°  87 BRCA2-11-F DNA Homo sapiens TTGGGACAATTCTGAGGAAAT SEQ ID NO°  88 BRCA2-11-R DNA Homo sapiens TGCAGGTTTTGTTAAGAGTTTCA SEQ ID NO°  89 BRCA2-12-F DNA Homo sapiens TGGCAAATGACTGCATTAGG SEQ ID NO°  90 BRCA2-12-R DNA Homo sapiens TCTTGAAGGCAAACTCTTCCA SEQ ID NO°  91 BRCA2-13-F DNA Homo sapiens GGAATTGTTGAAGTCACTGAGTTGT SEQ ID NO°  92 BRCA2-13-R DNA Homo sapiens ACCACCAAAGGGGGAAAAC SEQ ID NO°  93 BRCA2-14-F DNA Homo sapiens CAAGTCTTCAGAATGCCAGAGA SEQ ID NO°  94 BRCA2-14-R DNA Homo sapiens TAAACCCCAGGACAAACAGC SEQ ID NO°  95 BRCA2-15-F DNA Homo sapiens GGCTGTTTGTTGAGGAGAGG SEQ ID NO°  96 BRCA2-15-R DNA Homo sapiens GAAACCAGGAAATGGGGTTT SEQ ID NO°  97 BRCA2-18-F DNA Homo sapiens TGTTAGGGAGGAAGGAGCAA SEQ ID NO°  98 BRCA2-18-R DNA Homo sapiens GGATGTAACTTGTTACCCTTGAAA SEQ ID NO°  99 BRCA2-19-F DNA Homo sapiens TCAATAGCATGAATCTGTTGTGAA SEQ ID NO° 100 BRCA2-19-R DNA Homo sapiens GAGGTCTGCCACAAGTTTCC SEQ ID NO° 101 BRCA2-20-F DNA Homo sapiens GGCCCACTGGAGGTTTAAT SEQ ID NO° 102 BRCA2-20-R DNA Homo sapiens TTCCTTTCAATTTGTACAGAAACC SEQ ID NO° 103 BRCA2-21-F DNA Homo sapiens TGAATCAATGTGTGTGTGCAT SEQ ID NO° 104 BRCA2-21-R DNA Homo sapiens GTGTAGGGTCCAGCCCTATG SEQ ID NO° 105 BRCA2-22a-F DNA Homo sapiens CTGAGGCTAGGAAAGCTGGA SEQ ID NO° 106 BRCA2-22a-R DNA Homo sapiens CTGAGGCTAGGAAAGCTGGA SEQ ID NO° 107 BRCA2-22b-F DNA Homo sapiens GGTTTATCCCAGGATAGAATGG SEQ ID NO° 108 BRCA2-22b-R DNA Homo sapiens AGAAAATGTGGGGTGTAAACAG SEQ ID NO° 109 BRCA2-25-F DNA Homo sapiens CAGCAAACTTCAGCCATTGA SEQ ID NO° 110 BRCA2-25-R DNA Homo sapiens GGGACATGGCAACCAAATAC SEQ ID NO° 111 BRCA2-26-F DNA Homo sapiens GCACTTTCACGTCCTTTGGT SEQ ID NO° 112 BRCA2-26-R DNA Homo sapiens CGTCGTATTCAGGAGCCATT SEQ ID NO° 113 BRCA2-27-F DNA Homo sapiens CCCAGCTGGCAAACTTTTT SEQ ID NO° 114 BRCA2-27-R DNA Homo sapiens TCGGAGGTAATTCCCATGAC SEQ ID NO° 115 BRCA2-28a-F DNA Homo sapiens TCAAGAGCCATGCTGACATC SEQ ID NO° 116 BRCA2-28a-R DNA Homo sapiens AGGTAGGGTGGGGAAGAAGA SEQ ID NO° 117 BRCA2-29-F DNA Homo sapiens TGAGTCTACTTTGCCCATAGAGG SEQ ID NO° 118 BRCA2-29-R DNA Homo sapiens TTTTGCTTTCGGGAGCTTTA SEQ ID NO° 119 BRCA2-30-F DNA Homo sapiens TTTTTGCCTGCTTCATCCTC SEQ ID NO° 120 BRCA2-30-R DNA Homo sapiens GGTTTTTAAACCTGCACATGAA SEQ ID NO° 121 BRCA2-31-F DNA Homo sapiens TGAAATTTTGTTATGTGGTGCAT SEQ ID NO° 122 BRCA2-31-R DNA Homo sapiens TTTGAAATCTGTGGAGGTCTAGC SEQ ID NO° 123 BRCA2-32-F DNA Homo sapiens GTACCAAGGGTGGCAGAAAG SEQ ID NO° 124 BRCA2-32-R DNA Homo sapiens ATGGTGTTGGTTGGGTAGGA SEQ ID NO° 125 BRCA1-SYNT1-F DNA Homo sapiens TTCAGAAAATACATCACCCAAGTTC SEQ ID NO° 126 BRCA1-SYNT1-R DNA Homo sapiens TACCATTGCCTCTTACCCACAA SEQ ID NO° 127 BRCA1-S3Big-F DNA Homo sapiens AACCTTGATTAACACTTGAGCTATTTT SEQ ID NO° 128 BRCA1-S3Big-R DNA Homo sapiens CATGGGCATTAATTGCATGA SEQ ID NO° 129 BRCA1- DNA Homo sapiens CCTGCATGCTCATAATGCTAGA SExon21-F SEQ ID NO° 130 BRCA1- DNA Homo sapiens TTGGGATGGGTTTGAAGAGA SExon21-R BRCA1-1A DNA Homo sapiens SEQ ID NO° 131 GGGACGGAAAGCTATGATGTCACCACCGTCCGGGTGGGTGTGCTGGGGTTCACCCTCCCATTTCCC CAAGACCCCCTGCCAGGACATAGGCGGACGCGGGAGAGAAAACCAAAGAGGCTCCCTCCTTCCCCT TAGCATCTCTCTCCCGCCGTGTTCAGGAAGTGGATGGCTGCCCCAGCTCTTGTCCGCACTGGTACA CCTGCGTGCACGCGTGGGTACACAGCAGGCCCGAGCTTCGCGCTTGTGCCGCTCATATTCTACCCC TAAGAACTTCGCTTGAACTCTGACCTGCCCTTATATCCGAGAAAGTCAAATAAGCCCAGTTCGGCC TGTCCCAAACCGGCAGGGGCCCCTCAGACCACACCGGCGGGCTGGACCCCGGCTCTGAGGCCTCTG TTCCCAGGGCTCCGCCCAGATCTTCTGGGCCCCGCCCCCCGGCTGCGGGGGTGGGAGGAGGGGCCG GGGGGGCGCGGCCGCCTGGCTGGGGGCGGGGCGGAGGGGGGGCCGCGGACCCGGGGCGGGGGCTCG GCGCGGGCCCGCGAGATGCCGGTGTTGGCGGCCCGAGCGGCTGCAGTTGCAGGGGCGGGGGAGGCG GCGGCGGGGCCCGGGAGAGGGGTGGCGTGGGGGACCGGCGCGTAGCCGGGACCATGGAGGGGCAGA GCGGCCGCTGCAAGATCGTGGTGGTGGGAGACGCAGAGTGCGGCAAGACGGCGCTGCTGCAGGTGT TCGCCAAGGACGCCTATCCCGGGGTGAGGGACCTGCGTCTTGGGAGGGGGACGCTAAGGCTGCTGG GGGGTGGGTGACAGGGGCCCTGGCGACGGATGGGAATGGGTACTCGGGTAACCAGGGACAAGAGAC AGGGGGTCGGAGGACGCGGGGAGGCCTTGAGGGCTCAGGAAGGACTGCAGAGGATTGGGGTGGGAG GAATTAGGGAGCAGGGTGAGATAGATGGGGTTTGGGAGAACCAGAGCATCCGGGAGGGAGGGCGAG GGGAATGTCGGAGGTCCTGGGCAATGGAGAGGGGAAGAACTAGGGGGCTGAAGGGACCAGAAGGGA ACAGGAGGAGGTCTGGGAGCTTAGCAGAGATTCTCCGGGGGGGGGGGGGGGGGGGCAGGAGCTCCC GGGATCTCCCCTTTGCCCAATCCCAGACCAACTTGTGTCCAGGGGCTGGGCTGGACGGGGTGTGGG AGTGAGGAGGGCATTTATCTGGGGTGAGGACTTGGAGAGATGATCTCATCTGGATCCATCCGTGTC TGCAGAGTTATGTCCCCACCGTGTTTGAGAACTACACTGCGAGCTTTGAGATCGACAAGCGCCGCA TTGAGCTCAACATGTGGGACACTTCAGGTAGCCAAGTCCCTGGGGGTCACCCTGACTTCCAAGGCG GCCCACTCTGTCCCCTCCCTTGGTTAGACCCTTAGGTTCCAGGTAAGCCCAGCCCATCCATCCAAT TCCAACAGGAAGGGAAAAATCAATATTCTGCTAAAATCCAGGGAAACTGAGGTAGAACTTGCAGAG CCTGACAGAAACCATGTCCTGAAGGAGAAAGCCTAGGATCTGAGCCCCTCAGCTGGGTCCTGCCTA CCTGGGAAAGTTGGGAAGGAATGGCTTTTAATTTGGAACATGTTCCTTCAGAGATAAGACTGGGTT TAGAAAAGACATTTAGAGGCCAGGCACGGTGGCTCACGCCTGTAATCCTAGCACTTTGGGAGGCTG GGGTGGGGGGATCACCTGAGGTCAGGAGTTTGAGACCAGCCTGGCCAACATGGTTGAAACTCCGTC TCTACTAAAAATACAAAAATTAATCGGGCGTGTGGCACGTGCCTGTAATCTCAGCTACCAGGAGGC TGAGGCAAGAGAATCGCTGGAACCTGGGAGGCGGAGGCTGCAGTGAGCCGAGATCATGCCGCTGCA CTCCAGCCTGAGCGATAGAGCGAGACTCCATCTCAAAAAATAAAAAAGCAGAAAAGACATTTAGAA TGTCTTGAGTGAGGGGTGGTCAGGAGGCTGTTTCTCTCCATTGAACTAGATAAATCTGAGGTCAAG TCCCAGGAGAATGGGAGAGTGCTCTCCCTGCCACTGCTCTTTTCCTCCTCCCAACATAAGGAGGGT TTTTATTTTTACAAGAGTTCCCTTCAGGGCTTTAGACTGCCAAAGCCCAGAAAGCACATGCAACAT TTTATGAGAATGTCTATAGATTTTATGAGCTTCTCAAAGGGGTCCAAACCTCAGTCAAGAATAAAA ATTATTACTTTTTAAACCACTAGGGAAGCAGAGAGCCGTTTCCCACCATGTGACCTCCCTTCTGCC CGCTCCCCCACTTGGGAAACCCAGACTCCATGATGGGTATTAATGATGGGTATTAATGGTTGCTCT TTTCCATTCTCTGCTCCCAGCATCCCTTGACCAGGATCTGTAAGGTCTCCCATTCCCTTCCAGGCC TCCCATCCACTCAGGCCCCTCATGCCCTGTCTTCCTTCAGGTTCCTCTTACTATGATAATGTCCGG CCTCTGGCCTATCCTGATTCTGATGCTGTGCTCATCTGCTTCGACATTAGCCGACCAGAAACACTG GACAGTGTTCTCAAGAAGGTGGGAGCCTGGGGAAATAGGGCAGCTAGACTGAGGGGGACCAGACCA CCATGGTCCTGACATAACATGGGCCAGGAGGAGGGAGTGATGGCTGGGGTATGGCCATCAGCTGGT TAGCGAGTGAAGCTCTCATCCCTGCCACCCCTGCCTCCAGCCCCCATCCCTCCCAGCCACCCCTTT CCTGAAAGTCCTCAGAGCTGGATACAGCAGCTAGGGGAGGTGGGGGAGTGAAGGGAGAAGCACTCA CAGGATTCCTTCTCTGCTCTTCCAACTCCTTGGCAGTGGGAGTCCCAGATGGAGGGGATGGGATGG GAAGCCTGATCCTGGAGCTCAGGAAAGCCCTGTGGCCTCCTCTCCAGGCCCCAGTTTCCATGACAA AAGCCAGGGGTGAATGGACAGAAGTCAGCTAGGGCAGCCCCAGTTCCCAGGTGGGGGAGGGGAGGG TGGGATAAATTTGTTCCCAGGAGAGAGTATGGGAAAGGCGAGTGGGAATGGGAAGTTTCCAGGCTG GCAGACCCTTCATAGCCACTGAGGGAGAAGAGTCCACAGGCCCACGCCAGCCCTCTCCTCCCCGCT GCTTCTCTCTCACCCCATCCTGCTCTCAAACCAAGCCTAGCATTCTCACCTCCTTCCTCATGTGGG AGAGTCCTGAGGGATACATGGTTTCTGCGTGCTTGAGGAAGAGAGGGCACACTGCTGGCATGGCAC AAAGGCTCACGCTGTGCCTCCCTCCACCCCTCCACAATTCTCTTTTCTTCTCCTACATAGTGGCAA GGAGAGACTCAAGAGTTCTGCCCCAATGCCAAGGTTGTGCTGGTTGGCTGTAAACTGGACATGCGG ACTGACCTGGCCACACTGAGGGAGCTGTCCAAGCAGAGGCTTATCCCTGTTACACATGAGCAGGTG GGACCCTTGACGTCTGACCTCATCCCAGCCTAGACCTGTCACCTCTGCCC BRCA1-1B DNA Homo sapiens SEQ ID NO° 132 CCTCTGACCTGATCCCTTGACTGCCCCCAGCCTTGACATTCAACCCCAGCCCACAGCCTCCATGCC CCTTTCTAAGCTGCAGGCTAAGACCTATAACTTTCTCCCATGCACTCCTTCCTTTTCCAGGGCACT GTGCTGGCCAAGCAGGTGGGGGCTGTGTCCTATGTTGAGTGCTCCTCCCGGTCCTCTGAGCGCAGC GTCAGGGATGTCTTCCATGTGGCTACAGTGGCCTCCCTTGGCCGTGGCCATAGGCAGCTGCGCCGA ACTGACTCACGCCGGGGAATGCAGCGATCCGCTCAGCTGTCAGGACGGCCAGACCGGGGGAATGAG GGCGAGATACACAAGGATCGAGCCAAAAGCTGCAACCTCATGTGAGGGGCTAGGAGAGGGCAGAGT GTGAAGAGGGGTGGTGAGGGACACAATTGTTCCCCTGCCTGCGCCCAGGCTTCCTGACCTCCTGAT CCTGGCTGGGAAGTTAGGGCAGGCAGAGCGAGCAATTCTGGGCAGGGGAGCTGGAGGGCAGAAGGG TATCATCGTTTCTCATCTCCTCCTCCCTCCTCTTCTCCAGTGGATGTTGAGGGAGCTAACAGGGCT GGCATCTGGGGCATGAACTGGGATGGGGCAGGTGGGCGTTAGGGAAGCTGGTATCAAATGGTGACC TTGGTGGAGTCTCCTATGTGAAGAGTACCCTCCCTCTCCACCCCCAGTCCCCATATCCTGGTTCTG GCCCAAGGAAAATGTCCATTCTATGACCTTCTCTTTTCCTCTCCTCTCACTTCTGCAGCTATTCTC ACACATCTAACCTCTAGGCAACATGCACTAAATTCAAAAGCAAGGAGAAGCCCTTGCCCCCCATCA GTCCACCAGCCCTAGAACCTCCCTTGCCTCAACAGTCACCTAATAAAGCCCACCTCCATGGAAAAC GGCTGTGGCTTTAGTTTTGTTGCTTTTTAAAAAAATCAATCTACCAATCTTTAGCAGTAAGAGGGA AAGTTAGACCTCAGCTGGGGAACTTTCCTGTCCATGTCCACAGATAGAGCAGAGGACAAAGCCATA GGTTGGATCAGAAGTGTCCTTTTAGGAGTCAGAGTTGGGAGAAGGAGACATCCTGGGACTGTTCAT CCTAGTTAATGAAGTGGGCAATTCTCAGGCCATTAGGGGGTTTTAGAGCAGACCGACATATAATTA GTCAGCATTTCTCAGCCCAGCCAGGCCTGCTGCTAGTGTGGGAGGGGTCCTGCTCACCATCTGTAC CCCTGGCTTGGAGCCTGCTGGTACCCTGGGGGTTGTGGGGATAAGGAGGCATCAGGCCGGGCGCGC TGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGTGGGCGGATCACAAGTTCAGGAGATC GAGACCATCCTGGCTAACACGGTGAAACCCCATCTCTACTAAAAATACAAAAAATTAGCCAGGCGC GGTGGCAGTGCCTGTAGTCCCAGCTGCTCGGGAGGCTGAGGCAGGAGAATGGTGTGAACCCGGGTG AACCTGGGAGGCGGAGCTTGCAGTGAGCCGAGATTGCGCCACTGCATTCTAGCCTGGATGACAGAG CAAGACTCTGTCTCCAAAAAAAAAAAAAAAAAAAAAAAAGAAGGCATCAAAAGCCTCCACATCACA GAAGCTACCCCTGTACAGCGTGAAGTTTCCTAAGAGGTCAGTAGTTTGATTCTGGGGTCTCCTTAG AGGCTCAGGCCAGGGACCTTTCTCTCCTCCCATGCTGAGTTTCATGATGGCTTTCAGGGGAGCATC AGCTGTTAGAGTCACCCCTACCCTGTCCCTTAAAGGAAAGACGGTGGAGAGGACGGCTGAGCGCCT GTTGTCAGGAAAGACAGTACTGGTCTGTTTTCTCGGGAGTCTGGTTTCAGATTGTCCTGTATTCCC TTCCTGGCTCTGGTCCCACTGGCCTCTTTTCGGTGACATTCTCCCCCAGGAACCATCCCTGGCCCT TCCCTCCCCCAGCCCTAGCCAGTTCTCCCAGACACACTGGAAGAGAACACTGACCTTACCCAACTA TCTGCTGGGATCCCACCCAAATTTATAGCCCATTCCTCCCTCATTCATTCATTCAGCAAGTATGTA CTGAACACCAACTGTGTGGCATACACTGGCTTGGGAGATTGCAAGGACCAGTCTCTAAGCTTTTGG AGGCCAGCCCAGTGTGGAAGAGAGGTACCTCAGGTGTGAGGGTGCCATGGCTGAGGGATATTTGTA CATGTATGGGATGCTATGGGAGCTCCTTGCAGCCTGAGAAGCCAGTCCTGTGAGCCAGGTCCTGAG GGTTGAAGAGGAGTTTTCCGGGCAGGGAAGGGGTAGGAAAGGCACTCTGGGCAGAGGGTACAGCAT GTGTAAACACGTGGAGATGAGAATGAGCATAGCACTGTTGGGGCTCCCATGGCAGGGAGAATAGAA GACAAGGCTAGGAAGGTACACTGAGGCTACTGCAGGGTCCACAGAGGAATCAGAATTTCATTCTGA GGATGAATGAAATCATCCTCAGAGGATGAAGCCACCAGGAATTTCAGGCAGAGAGTGAAGTGATCA GAGTTGTTTTTTGGATAGATGGTTATCTGGATGTGGTGTTGGAGCTGGGAGATTTGGCTCTGAGGT GTGTCATTTAAAATAATAGCTTCTCGGCAGTGGCTCACACCTATAATCCCAGCCAAGATTCCTCCT TTGGGAGGCCAAGCTGGGAGGATCGCTTGAGGCCAGGAGTTAGAGACTGCAGTGAGCTATGATCAT GCCATTGTCTTCCAGCCTGAGTGTCAGAGTGAGACCCTGTCTCTAAAAAAAATTAAAAAATAAAAA ATAAAAAATAGCTTCTCCTTTCCCTTATGCCAGGTTCCAGTCTTGAGAGGAAAGGAATCCCTACCC ACCACTCCCTGGATCATCAGATATCCCTATCCCAACCTCTCCTATGGGACTAGTTCATCTCAGCCA GTCTCAAAGATTCTAGGATAACTTCAATGGCATTTGAAATTATCTAAGTGTGCTTGGATAACCACC CCCTCAAACTGAGACCTGGTTAGGGACTGACTCAAAGACCCTGAGTCCTCGGCTAAGGGTACAGGA GAGGGCAGGGGCTCCAGGCCCAGCTAGGTGGATCTCCATCTGTCTCTGAGGACTGACCCTTTCCCC ACAAGGACCTGCCATAAAAATCGACTTGCGATTTTTAGCTGAGTGGCTTCTCTTTTCCACTTTGGA CTTCTCAGTGTATAGCAGGTTCAAGCCTGCAACCACCAAAGTGCAGAGTGTGGAGTGTTTGTGCCC CCTCTTTCCTCCAACCTCCATATCCTGCCATGTGAGCTCAGGGAATGCAAATGCATTTAAATATCC ATCTAAAGCAAACATAATTAGAAAAATCAATCAGCTGGAGGACCCCCCAAAGTTTAATACATTTTC AATACCACCAGGAATGGATTTTTGGTCCCTTTCTGCAGGTCTGGGTTGCCAGACGTTTTATTTCTG GGGAGGAGGGCTCTGGGCTGAGGAGCTCAGTGGGTGGGAGGAGGGAATGGGACTGTTGCTGAT BRCA1-SYNT1 DNA Homo sapiens SEQ ID NO° 133 TTCAGAAAATACATCACCCAAGTTCCCATCCCTACCTGTCTATCCACAAAACCAAGGCATTCCTGA GATTAGTTCATTTATTATACTAATATAACAAGTGTTTATTAAGTATCTACTACTATATTCAAGTAC TATTCTAGGAGATAGAAATGTAGCAGTTTACAAAATAAAGCCTGCTCTCATAGAGCTCATATTCTA GTGTGGTAGACAGTTGATACGGAATTAAAGAATACATGGGAATAAGTGCATTAAAGAGAAAAATTA AGCAGGGTAAGGGGAAACAGGTAGTTCAATATCTATGTGGGGGTGAGATGTACATGGGGGGAGTCA GGAAAGGTTTCACTGAGGTGAGACTAGAGGATAGCTTAATAATGTAAAGAAACACACTATGCAACA ATTAGGGGAAGAGCATTCCAAGAAAGAGGGAGCAGAGAAGGCAAACCCTGAGCAGGACCATGCCTG TGTATGCAGGACATCAGATAGGTCAAGGTGCTAAAATGTAATAATCCAGGAGGATATTGTAGGGAA AGACTATCAGAGAGGTAGCTGGTAACTTCTGGTAGGAACCTATAGGCTATTTTAAATCTTTAGCTT TATTCTGGTCTTTTTAATTTTCTTTTTTTTTTTCAGACAGAGTCTCGTTCTGTCGCCCAGGCTGGA GTGCAGTGGCACCATCTCGGCTCTCTGTAACCTCCGCCTCCTGAATTCAAGTGATTCTCCTGCCTC AGCCTCCCGAGTAGCTGGGACTAAAGGCATGCACCACCATGCCTTGGCCTCCCAAAGTACTGGGAT TACAGGAGTGAGCCACCATGCCAGCCATCTTTTTAATTTTTAATGTTAATTAATTTTTGTAGAGAC AGGATCTCACTATGATGCCCATGCTGGTCTTGAATGCCTGGCATCAAGCAATCTTCCTGCTTCGGC TTCCCAAAGTGCTGGGATTACAGGTGTGAGCTACTATACCCGGCCTTTAGCTTTCTTCTGAATGTG AACCTTTTTTTTTTTTTTTGGAGATGGAGTCTCACTCACTCTGCTGCTCAGGCTGGAGTGCAGTGG TGTGGTCTTGGCTCACTGCAACCTCTGCCTCTCGGATTGAAGTGATTCTTGTGCCTCAGCATTCCA AGTAGCTGGGACTACAGGCGCGTGCTGCCACACCCGGCTAATTTTTTTGTATTTTTGGTAGGGAAG GGGTTTCACCATATTGCCCAGGCTGGTCTTGAAGTCCTGACCTCAAGTGATCCATCTGCCTCGACC GGGATTACAGGCGTGAGCCACTACACTTAGCTCTAAATGTGAATTTTTGAAACGGATTTTTTGGAT AAAGTCCAGGCAAGATATCAAAGAACGACTAACCTGGCAGTGTGACAAGAATGTGGTTTTTTCCTT AAATATTTAACTTTTTAGAAAAGGATCACAAGGGCCAGGTGCGGTGGCTCACGCTGTAATCCCAGC ATTTTGGGAGGCCAAGGCGGGCCAGCCTGGGTGACAGAGAATCCATCTCAAAAAAAGAAAAAAAAA AAAGAAAAGGATCACAAGAAAAGCTTGTGGACAGTAACCTTATTGTGAAGGGTTGTAATACAACTC TTGTAATCATGGGGTTTTTGACATAGCACAGGGCAGTGAAAAGAAAAACAATGAACTAAGTCAGGA GGCTGGGTTTCTACTACCAGTTGTGTATATAAGCAGAGCCACCTTGGGCTAACCACTCTACCTGAA CCTGTTTCCTTCTCTTGCCATTCACCCTGCCAGACTCCTTGGGCTATTGCAAGAATAAAATTAAAT GCTACTTGGGAAAATGCTTCACAACCTGAGATGACTTGGGAAAAATGCTTCACAACCTGAGATAAC TTGTACCAACATTGGTATTATTACTGGGACCAAATGTGACTTTAAAAAGAAAAACAACCTTGACAA AGAAAACTCTGATTGGTTACTAAATCCCTATTTCTGAGATAAGCTACATTTCAAAGAAATTCTCCG TAAAAGAAAAATTGGATTCAGTTATCATACCAGATGGCTTTCATTCTCACCACTGACTCAATTCTG AAACAATTATATTTCAGTATGGTAATTATAATCTAAACTATATAAACACACTGTAAACACAAACTT TGAACAGATGAAAACTCCGATATGTAAAAAGGTAATGAATGTTGAAGGAAGACTGTGAAAAGGGAA AAGAAAAAAAATTAAAATGTTCCCCTTCTAGGTCCTGATGAGAGTAAATGTTTACTATAAAAATGA TTCAAATATTTTAAACACTTTTCAAACCAGGCAATATTTTAGGCCTACTGTATATTTGCATTTTGA GCTTCCAATACGGATAAGTGACTGGAAAAAGCAGCTAGGTTTAGGTTGAAAAACAACAACCCACCG GGGAACACATTTTAGCAAATTCTTCTGAAAGTCAAAAATGTTATAGTCATAGGTAAAAAGTTACAA AGAACTACCAATTGTCAGAAATAGCTGCCAATATTGACTTAGAAGACAGCAGAAGGAATTTTAGTT CAAGAAACCTAAAACAGGCTGAAAACCTTACCTACCCTATAGCTACCACAAATAACACTGTTTCCA GTCATGATCATTCCTGATCACATATTAAGACATAACTGCAAATTGTGCTATACTGTACTATATTAA AAGGAAGTGAAATATGATCCCTATCCTAGAACTTTCCATACAAATGAATGTAAAACACCATAAAAA TTAATCTTAAGGCCGGGCGCGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGTGGG CGGATCACGAGGTCAGGAAGTGGAGACCATCCTGGCTAACACGGTGAAACCCCGTCTCTACTAAAA ATACAAAAAATTAGCCGGGCGTGGTGGTGGACGCCTGTAGTCCCAGCTACTTGGGGGGCCGAGGCA GGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATGGCGCCACTGCACTCCGGC CTGGGTGAAAGAGCGAGACTCCGTCTCAAAAACAAAACAAACAAAAATTAATCTTAAGCCAGGCGC AGTGGCTCACGCCAGCACTTTGGAAGGCCGAGGCGGGTGGATCACGAGATCAGGACTTCAAGACCA GCCTGACCAACGTGATGAAACCCTATCTCTACTAAAAATACAAAATTAGCCGGCCACGGTGGCGTG CGCCTATAATCCCAGCTACTCAGGAGGCTGAGGCAGGAGAAGCGCTTGAACTTGAACCTGGCAGGC GGAGGTTGCAGTGAGCCAAGATGGCGCCACTGCACTCCAGCCTGGGCGACAGAGCCAGACTCCAAC CCCCCACCCCGAAAAAAAAAGGTCCAGGCCGGGCGCAGTGGCTCAGGACTGTAATCCCAGCACTTT GGAAGGCTGAGGCGGGTGGATCACAAGGTCAGGAGATCGAGACCATCTTGGCTAACATGGTGAAAC CCCGTCTCTACTAAAAATACAAAAAATTAGCCGGGCATAGTGGTGGGCGCCTGTAGTCCCAGCTAC TCGGGAGGCTGAGGCAGGAGAATGGCCTGAACCCGGGAGGCGGAGCTGGCAGTGAGCCAAGATCGT GCCACTGCACTCCAGCCTAGGCAGCAGAGCGAGACCGTGTCTCAAAAAAACAAAACAAAACAAAAC AAAAAGTCTGGGAGCGGTGGCTCACGCCTGTAATCCCAGCACTTTCGGAGGCCAAGGCAGGAGGAT CACCTGAGGTCAGGAGTTCGAGACCAACCTGACCAATATGGAGAAACCCTGTCTCTACTAAAAATA CAAAATTAGCTGGTGTGATGGCACATGCCTGCAATCCCAGGTACTCCGGAGGCTGAGGCAGCAGAA TTGCTTGAACCCGGGAGGTGGAGGTTGTAGTGAGCCGAGATTGTGCCACTGCACTCCAGCCTGGGC AACAAGAGCCAAAGTCTGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAGAAATTAATCTTAACAGGA AACAGAAAAAAGCAATGAAAAGCTAGAAAACATAATAGTTGATTGAAAATAACAATTTAGCATTTT CATTCTTACATCTTTAATTTTTATGTATCTGAGTTTTTAATTGATGGTTTAATTTGCCAGAATGAG AAAGAACATCCTATTTTTATGACTCTCTCCCATGGAAATGAAACATAAATGTATCCAAATGCCACA CTATTGAGGATTTTCCTGATCACTGATTGTCATGAGTAAGTTTTGTGCTTTTTCAAAAGCAGTTTT TTCCTACAATGTCATTTCCTGCTTCTCTGGCTCTGATTTTCAATAAATTGATAAATTGTGAATCCT GTTTTCCTCTTATTTTTGTTTAGCTATAATGTTGAAGGGCAAGGGAGAGGATGGTTATTTATAAAT CTTGTATCGCTCTGAAAACACAACATACATTTTCCTTAATCTGATTAACTTGACTTCAAATATGAA AAACAACTTTCATAAAGCAGAAAAGAATTTACCCTTTTTTATTGTGGGTAAGAGGCAATGGTA SEQ ID NO° 134 ForwardPrimerPrefix DNA Artificial Sequence AAAAGGCGCGCC SEQ ID NO° 135 ReversePrimerPrefix DNA Artificial Sequence AAAATTAATTAA

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RELATED PATENTS AND PATENT APPLICATIONS

-   Lebofsky R, Walrafen P, Bensimon A: Genomic Morse Code U.S. Pat. No.     7,985,542 B2 (application Ser. No. 11/516,673) -   Murphy P D, Allen A C, Alvares C P, Critz B S, Olson S J, Schelter D     B, Zeng B: Coding sequences of the human BRCA1 gene U.S. Pat. No.     5,750,400 -   Skolnick M H, Goldgar D E, Miki Y, Swenson J, Kamb A, Harshman K D,     Shattuck-eidens D M, Tavtigian S V, Wiseman R W, Futreal A P:     17q-linked breast and ovarian cancer susceptibility gene U.S. Pat.     No. 5,710,001 

1. A composition comprising at least two polynucleotides wherein each polynucleotide binds to a portion of the genome containing a BRCA1 and/or BRCA2 gene, wherein each of said at least two polynucleotides contains at least 200 contiguous nucleotides and contains less than 10% of Alu repetitive nucleotidic sequences.
 2. The composition of claim 1, wherein said at least two polynucleotides bind to a portion of the genome containing BRCA1.
 3. The composition of claim 1, wherein said at least two polynucleotides bind to a portion of the genome containing BRCA2.
 4. The composition of claim 1, wherein each of said at least two polynucleotides contains at least 500 up to 6000 contiguous nucleotides and contains less than 10% of Alu repetitive nucleotidic sequences.
 5. The composition of claim 1, wherein the at least two polynucleotides are each tagged with a detectable label or marker.
 6. The composition of claim 1, comprising at least two polynucleotides that are each tagged with a different detectable label or marker.
 7. The composition of claim 1, comprising at least three polynucleotides that are each tagged with a different detectable label or marker.
 8. The composition of claim 1, comprising at least four polynucleotides that are each tagged with a different detectable label or marker.
 9. The composition of claim 1, comprising three to ten polynucleotides that are each independently tagged with the same or different visually detectable markers.
 10. The composition of claim 1, comprising eleven to twenty polynucleotides that are each independently tagged with the same or different visually detectable markers.
 11. The composition of claim 1, comprising at least two polynucleotides each tagged with one of at least two different detectable labels or markers.
 12. A method for detecting a duplication, deletion, inversion, insertion, translocation or large rearrangement in a BRCA1 or BRCA2 locus, BRCA1 or BRCA gene, BRCA1 or BRCA flanking sequence or intron, comprising: (i) isolating a DNA sample, (ii) molecularly combing said sample, (iii) contacting the molecularly combed DNA with the composition of claim 5 as a probe for a time and under conditions sufficient for hybridization to occur, (iv) visualizing the hybridization of the composition of claim 5 to the DNA sample, and (v) comparing said visualization with that obtain from a control sample of a normal or standard BRCA1 or BRCA2 locus, BRCA1 or BRCA gene, BRCA1 or BRCA flanking sequence or intron that does not contain a rearrangement or mutation.
 13. The method of claim 12, wherein said probe is selected to detect a rearrangement or mutation of more than 1.5 kb.
 14. The method of claim 12, further comprising predicting or assessing a predisposition to ovarian or breast cancer based on the kind of genetic rearrangement or mutation detected in a coding or noncoding BRCA1 or BRCA 2 locus sequence.
 15. The method of claim 12, further comprising determining the sensitivity of a subject to a therapeutic treatment based on the kind of genetic rearrangement or mutation detected in a coding or noncoding BRCA1 or BRCA 2 locus sequence.
 16. A kit for detecting a duplication, deletion, inversion, insertion, translocation or large rearrangement in a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron comprising a) at least two polynucleotides wherein each polynucleotide binds to a portion of the genome containing a BRCA1 or BRCA2 gene, wherein each of said at least two polynucleotides contains at least 200 contiguous nucleotides and is free of repetitive nucleotidic sequences, wherein said at least two polynucleotides are tagged with visually detectable markers and are selected to identify a duplication, deletion, inversion, insertion, translocation or large rearrangement in a particular segment of a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron, and optionally, b) a standard describing a hybridization profile for a subject not having a duplication, deletion, inversion, insertion, translocation or large rearrangement in a BRCA1 or BRCA2 locus, BRCA1 or BRCA gene, BRCA1 or BRCA flanking sequence or intron; c) one or more elements necessary to perform Molecular Combing, d) instructions for use, and/or e) packaging materials.
 17. The kit of claim 16, wherein said at least two polynucleotides are selected to identify a duplication, deletion, inversion, insertion, translocation or large rearrangement in a particular segment of a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron associated with ovarian cancer or breast cancer.
 18. The kit of claim 16, wherein said at least two polynucleotides are selected to identify a duplication, deletion, inversion, insertion, translocation or large rearrangement in a particular segment of a BRCA1 or BRCA2 locus, BRCA1 or BRCA2 gene, BRCA1 or BRCA2 flanking sequence or intron associated with a kind of ovarian cancer or breast cancer sensitive to a particular therapeutic agent, drug or procedure.
 19. A method for in vitro detecting in a sample containing genomic DNA, a repeat array of multiple tandem copies of a repeat unit consisting of genomic sequence spanning the 5′ end of the BRCA1 gene wherein said repeat array consists of at least three copies of the repeat unit and said method comprises: providing conditions enabling hybridization of a first primer with the 5′ end of the target genomic sequence and hybridization of a second primer with the 3′ end of said target sequence, in order to enable polymerization by PCR starting from said primers; amplifying the sequences hybridized with the primers; detecting, in particular with a probe, the amplicons thereby obtained and determining their size or their content, in particular their nucleotide sequence.
 20. The method of claim 19 wherein the repeat unit encompasses the exons 1a, 1b and 2 of the BRCA1 gene and optionally encompasses a sequence of the 5′ end of the NBR2 gene.
 21. The method of claim 19, wherein the downstream and upstream primers are respectively selected from the group of: for a downstream primer: a polynucleotide sequence in the region between exons 2 and 3 of BRCA1, preferably at a distance from 2-4 kb from the 3′ end of exon 2, more preferably at a distance from 2.5-3 kb from the 3′ end of exon 2 or a polynucleotide sequence in the region between exons 2 and 3 of BRCA1, within 2 kb from the 3′ end of exon 2, preferably within 1.5 kb and more preferably within 1 kb from the 3′ end of exon 2 for an upstream primer: a polynucleotide sequence in the region between the BRCA1 gene and the NBR2 gene, within 2 kb from exon 1a of BRCA1, preferably within 1.5 kb and more preferably within 1 kb of exon 1a of BRCA1 or, a polynucleotide sequence within exon 1a of BRCA1 or within exon 1b or in the region between exons 1a and 1b or, a polynucleotide sequence in the region between exons 1b and 2, or in exon 2, or in the region between exons 2 and
 3. 22. The method of claim 19, wherein the primers are selected from the group of: BRCA1-A3A-F (SEQ ID 25), BRCA1-A3A-R (SEQ ID 26), BRCA1-Synt1-F (SEQ ID 125) and BRCA1-Synt1-R (SEQ ID 126) or their reverse complementary sequences. 